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I performed real-time PCR on cDNA of samples that had a nanodrop quantity of 1000+ ng/ul, but when i perform i do not get any proper results. i am attaching the report of my real-time. kindly help me out. should i synthesize the cDNA again or should i change something in the way i'm performing real-time pcr.
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Kübra Cansu Y. Yıldırım
when I had Put the pcr tubes in the thermocycler for cDNA synthesis, there was a power breakage of 5 minutes after which the reaction resumed. I performed the nanodrop which gave satisfactour results, the comcentration and purity of the cDNA were both good. But after performing real-time pcr, I’m not getting results. should I synthesise the cDNA again too or should I use the previously synthesised cDNA (used in this reaction) for real-time again?
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I do run for 2 sample real time pcr .. their melting curve for one sample show not available tm what this mean ??? Its ct value is 31
thanks alot
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Hello. If their first cDNA concentrations were equal for each sample, your second sample has a lower amount of final amplification. It depends on the efficiency of your primer and its TM. Did you optimize your primers? And what was the gene? I had seen that mutations could influence primer annealing and melting curve
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I used a KOD DNA polymerase for PCR that works very well. Then I used this enzyme to run a TaqMan probe real-time PCR but the results waere unexpected. I attached the results. The fluorescence rise very fast during initial cycles and it reach to a plateau much faster than the control. The control enzyme is a Hot-start but the KOD is not. Can this be the cause of this phenomenon or is there another reason involved?
figure legend:
Red: Control enzyme (Hot-start)
Blue, green, purple, and pink: KOD with different buffer.
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Hi
Yes, I also agree you need to optimize the reactions first, what ever polymerase you are using. Also the nature of enzyme (hot start/regular) will also make difference.
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HiL simualtions are now being promoted as fast and accurate validation of power electronics systems. For ex., I have used Plecs+RT box for validating the performance of some converters. However, I have a question, what are the practical relevance of the HiL tests ?? In other words, what is the added value of the HiL results compared to what could have been achieved with a detailed offline simulation ? (considering representation of all relevant delays, discrete time implementation of the control loops etc. as can be easily included in any offline simulation in similar ways as in the real-time simulation) #research #power #electronics
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The HIL setups allow us to test a real piece of hardware (hence the name - hardware-in-the-loop) with its pertaining features and operational modes.
Specifically, in a controller-hardware-in-the-loop (C-HIL, just one type of HIL setup) setup we can test how the controller with an embedded code is going to behave once deployed in an actual device. What is hard to model or do in e.g. MATLAB in this regard is the following:
- Controller capacities utilization - in various forms! (memory overrun, interrupts execution, etc.),
- Coordination and execution of different control layers (especially if they are executed on different controllers/control platforms),
- Communication devices and systems, overlaying the power electronics system (how are you going to model dynamics associated with the CAN communication protocol, for example, used to exchange the references between the controllers?),
- Integration with other real-time executed tools (for example, flight simulator for testing more electric airplanes),
- etc.
So to abstract things - Doing tests on a certain piece of hardware in real-time allows us to connect these devices to other devices and processes that are also executed in real-time (and sometimes can not be easily executed in non-real-time context) and to examine many features that are simply hard to model, or sometimes meaningless, in an offline simulation.
On the other hand, and more broadly speaking, unlike MATLAB, the HIL setups allow us to model and test:
- Medium/large complexity and size systems (for example, microgrids are extremely hard to model in detail since the model runtime tends to increase exponentially),
- High-power/high-voltage devices (in a power-hardware-in-the-loop, P-HIL, setup we can test the behavior of the real protection device in various network situations, such as short-circuits, loss of phase, etc. - this is next to impossible to do in offline simulation),
- etc.
This all being said, the offline simulation tools (constituting model-in-the-loop, MIL, setup) are here to stay. They are still great for initial tests, to put it in simple terms, and simulation tool providers are constantly adding new useful features, but the HIL setups have closed the serious gap that existed between the offline simulations and the tests on actual devices (prototypes or commercially available devices). It is safer than the real hardware, but faster and 'more precise' (again serious simplification) than the models executed in offline simulations.
You can check the following two documents for further details if you wish.
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hi , if you please i need to know what are the reasons of the false positive results in real time RT-PCR of non specific template reaction, the primers are not similar to the non specific template while the probe is close similar to the non specific template? could i get an explanation ?
Definitely, my problem is in the reactions that have a DNA but it is supposed to be non-specific for the primers and probe. After checking the primers and probe sequence, I found that the primers were completely non-specific for this DNA but the probe was specific partially. so, my question here is, can the reaction be falsely positive because of the probe?
thank you
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Do you mean that you blast the primers and probe sequences against the non specific target and you found a degree of matching between your probe and the non specific target?
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I want to measure the IFN-alpha subtypes produced by plasmacytoid dendritic cells..I think it can be measured by real time PCR, but I can't find the primers for subtypes of IFN-alpha. Can anyone guide me on how to go about this experiment? Or have experience in such quantification? Please help me out..
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You can find the specific sequence of IFN-alpha from NCBI database . Take the gene sequence make your primers pair using Primer Quest ( IDT).
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Can anyone suggest me a free plataform or API to retrieve data from sensors (in real time if possible)?
The data type can be about weather, agriculture or similar.
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In fact I only hope to retrieve real data (through an API, for example) to use data interpolation techniques.
I believe that there must be some API that allows me to retrieve data from sensors as close as possible to real time.
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I am trying to conceptualize a topic on radon monitoring in the Philippines and I am still thinking if using CR-39 detector is enough or should I incorporate the usage of the RAD7 real-time monitor for radon? Thank you.
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Mattia is correct. For a home, the CR-39 is for a minimum of 90 days and the RAD7 is for 1 hour house measurements over a few days. To find hot spots, use the RAD 7 (be aware of Rn-220 response)
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Hello,
I am working on lncRNAs, one of my lncRNAs is INHBA-AS1 and I design intron spanning primer for it. In real time PCR I just have primer dimer with melt lower than 70, I decreased primers concentration and increased cDNA concentration and also increased annealing tempreture but I just have primer dimer.this is my primer's image from NCBI. what should I do?
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thanks a lot Frederic Lepretre for answering my question, actually I work on placenta tissue and whole blood and I checked the expression in NCBI but I didn't check on GTEX.
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Investments are real-time updated science, due to its special properties. Therefore, tell us what the last updates and topics about it are.
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High-yield savings accounts.
Short-term certificates of deposit.
Short-term government bond funds.
Series I bonds.
Short-term corporate bond funds.
S&P 500 index funds.
Dividend stock funds.
Value stock funds.
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We assume that the most likely answer is unfortunately yes.
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Solving an equation in real time is about the speed with which it can be done. Also, the context would be important. Typically heat flows are relatively slow -- seconds rather than milli- or micro-seconds or less (as might be appropriate for various wave systems, esp. electromagnetic ones). Weather problems include heat/diffusion effects & equations, and need to be solved over hours to provide "real time" predictions.
That being said, if the problem is say essentially two-dimensional (plates, perhaps), then the solvers can be quite fast. Large 3-D problems with complex geometries would be harder, but could probably be done with some clever computational methods, such as
* precomputing the mass/stiffness matrices (perhaps using a lumped mass approximation),
* solving the equations coming from an (implicit) ODE solver for the finite element/difference/volume equations using iterative methods and good starting approximations (say, a low order predictor to feed into the implicit solver).
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Hi,
I have taken motor imagery EEG signals from two healthy persons from a hospital. can you suggest the details of documents to be submitted in a journal (in order to provide proof for the real-time analysis)? I have received a bonafide certificate(with doctor's sign and seal) stating that the recordings are taken under doctor's observation and it is eligible for doing experimental analysis. Is this proof enough for submitting in a journal?
Note: This analysis didn't involve patients with disorders
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Different journals will have different requirements for what you call proof. It is difficult to make one statement to cover all the journals so I suggest you may need to contact a journal that you are interested in and ask them what they need exactly if this is not clearly stated already on their website in "Instructions for Authors".
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With ongoing development of real-time "Free Style Libre" insulin detectors, has anyone ever did insulin analysis over a week period for a wild "forest" mice (not wild type lab mice)?
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No special clinical need or product measurement.
Thought it could have been a blank control in some metabolic intervention experiments.
There are no data on insulin levels in indigenous mice population within their normal food environment, rather than WT one produced in the lab.
Reason: similar comparison can be projected between two human populations.
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So here is the condition -
I have to quantify two genes in two different membranes, these are OCT4 and NANOG genes. These are to be quantified on two different grafts. I have RNA of the graft, cDNA is there also. But the primers that I am having have sizes as follows -
OCT4 - 393bp
NANOG - 75bp
These are RT PCR primers that are there in lab. Now as there is too much size different between the two, should I be using two GAPDH (as housekeeping) for them separately. That is in reaction one NANOG + GAPDH1 (with amplicon expected size 98) and in reaction two OCT4 + GAPDH2 (with amplicon size 380 bp). I have to quantify these genes between two tissue types. I am not interested to know expression of NANOG relative to OCT-4 in say graft 1 or graft 2, I want to now expression difference between NANOG expression in graft 1 and 2, and the same with OCT4 gene.
regards for your help.
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try to use 1 HKG and see how it works
if it gives you fantastic result
you are good to go
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hii,
I have one group in six different ZTs --> ZT0, ZT4, ZT8, ZT12, ZT16 e ZT20.
I performed gene expression by real time PCR using the 2- deltaCt method
I have used graphpad to represent the curve but I don't know what statistical method to apply to obtain the p value.
Thanks in advance :)
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Thank you for all your answers, I will try it. :)
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I want to read researches about safety critical systems in industry, and what I find are works related to transportation industry and medical industry.
Are there any other related researches in other branches of industry? like industry 4.0 or petroleum industry?
Also, I want to know if there are any researches related to safety of small units in industry
If there are, what are the best keywords to search with? and what are the best databases?
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One of the important areas of research for Industry 4.0 is human performance enhancement. You may find these articles relevant to your question and some of the references in the papers may also be helpful.
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I have been working on a Home Automation System in which several loads have been connected. The main controller is Esp32. The voltage and current sensors are connected with/across the loads and the values are sent to the controller. Now these values are visible in "Arduino Serial Monitor" in real-time. But I want to send these values to a real-time database (e.g. Firebase, InfluxDB etc.). Currently I have been stuck in exactly this phase. Kindly help me out and do reply me if anyone knows how to send these real-time values of voltage, current and power to a real-time database.
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The real time product was run on gel . What could be the reason ? Are the both bands dimer???
Or may be due to ladder problem??
The melt curve peak is about 85c.
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@ Paul Rutland
Thanks .
Yes all the samples should amplify the product .
I will check it and will share the results.
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I'm getting multiple peaks in my real time pcr melt curve for tnf alpha gene. Amplicon size is 284 .
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if your curves are on the left side, it may be primer dimers; on the right side, it may be genomic contamination. I think you can increase Tm in the cycles or decrease the primers concentration. and it would be better. BTW the way 284 is high but you can get the result.
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Hello everyone
I would like to know if someone have experience in the process of synthesis of primers and probes for a qPCR real time (in vitro), for this process, what type of material or machine i will need?, what environmental conditions i should have? , what type of professional will do this process? maybe a biotechnologist?
If someone have information about this, i really will be grateful.
Thanks a lot.
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Thank you very much
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I'm using the Tri-zol kit, and isolating using Isopropanol.
I've already tried increasing the number of Ethanol washes, and a second Chlorophorm step yet my 260/230 remains consistently under 0.5.
Does anyone have any ideas where the contamination is coming from?
I need the RNA for real time rt-PCR, can I use it anyway if my 260/280 is ok?
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For isopropanol precipitation use EQUAL volume. Follow the Trizol protocol. For 1 ml Trizol, add 200 ul chloroform to extract, mix well, spin at high speed and remove aqueous top phase, which should be 500-600 ul. Dont take any interface, no need to get every last drop. Add an equal volume of isopropanol and vortex. No need for 4oC overnight. Leave at room temp and spin at high speed , then wash pellet in 70% ethanol. Usually no need for DNAse if your primers for RT-PCR are in two separate exons.
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real time Eye movement tracking analysis
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Maybe these articles will help you.
1) Neuro-Inspired Eye Tracking With Eye Movement Dynamics (thecvf.com)
2) Best practices in eye tracking research - ScienceDirect
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is it possible to control non-linear changes with PID?
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Universal control channel https://t.me/universalcontrol
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I have been working on a Home Automation System in which several loads have been connected. The main controller is Esp32. The voltage and current sensors are connected with/across the loads and the values are sent to the controller. Now these values are visible in "Arduino Serial Monitor" in real-time. But I want to send these values to a real-time database (e.g. Firebase, InfluxDB etc.). How can I send these real time values of Voltage, power and current to the real time database?
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What about these?
BTW.: g..gl. is a great resource to answer such questions on your own.
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i have been developed a taq man real time pcr for my work… but i have a problem in maintaining my STANDARDs and after a week the load of plasmids gets low… how can i solve this problem?
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It may be that your dna is absorbing onto the plastic of the tube or it is being degraded by nucleases or acid pH.
You can store lyophilised and make up in TE just before the assay or add some non amplifying dna or rna to your standards before storage. The cold dna will block the absorption sites on the plastic tubes. Siliconisation of the storage tubes may also help
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I am using Step One real-time instrument and the Green master mix with ROX.
Thank you all
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Vipul Batra thank you, I will check it.
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Dear RG experts,
The statistical study includes a number of tests, some of which are well-known, while others are controversial. For me, applying such dubious standards to real-world problems is a major issue. I learn all of these in order to solve real-life problems.
Please help me with the application of the run test.
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Respected James R Knaub Sir,
Thank you so much. Further, I will ask about some problems with it.
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Our group is trying to pull down AGO-FLAG protein to look for miRNAs interacting with AGO2, however we are getting expression at the same Ct for non-specific miRNAs in the negative control group as well in the real time. How to improvise our technique to get good results ?
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hi,
Non-Specific binding (NSB) refers to an occurrence of an antibody binding to unintended proteins, receptors, or transporters. This binding of assay antibodies is not correlated with the specificity of the antibodies.
ips for Reducing Non-specific Staining in IHC
  1. Fixation. Over-fixation can cause high levels of background in IHC experiments. ...
  2. Slide Preparation. Decreasing the thickness of tissue sections can help keep reduce background staining. ...
  3. Deparaffinization. ...
  4. Blocking. ...
  5. Antibodies. ...
  6. Endogenous Peroxidases and Biotin. ...
  7. Further reading:
Strategies to reduce non-specific binding
  1. Adjusting the pH of your buffer.
  2. Using protein blocking additives.
  3. Adding non-ionic surfactants.
  4. Increasing salt concentration.
For more information kindly refer this link:
Best wishes..
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I am working on research topic "QoS mechanism in IoT". I have to implement QoS in Cooja simulator. My questions are:
1) How can I classify 3 types of applications (Critical, Real time, Non-real time) in cooja simulator? Which method of cooja can make these nodes different from each other?
2) How I can implement multiple queues in cooja (weighted fair queue) one queue for each type of traffic?
Can anyone help me in this implementation? or can implement it?
Thank You
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Question-7: Are our mathematical means ready for rigorously expressing software system requirements? Is our computer’s inference power adequate for expressing real-time inexhaustive, indeterministic, and uncertain behaviors in SSE?
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Hello
Dear researchers
In recent years self-balanced switched capacitor boost Multilevel inverters become a more trending research area. In which the capacitor is connected across the DC source to charge in a short period. But in the practical application how far this is feasible without the current limiting inductor.
1) Maybe in most cases, researchers are assuming the initial capacitor voltage is charged at its rated position. otherwise, it would draw a very high surge current from the input source in starting. In real-time there will be a voltage deterioration when it is not in operation. so if it is switched it will damage the IGBT?
2) I have observed in the MATLAB/ simulation. The switch resistance is considered as 1m.ohms(Ron). Even in normal conditions also the capacitor draws impulses current is nearly 100 times of rated load current. but that impulse is not observed in DSO(oscilloscope) while measuring the capacitor voltage in real-time experimentation may be due to IGBT resistance(it is approximately 14m.ohms).
4) what is the impact of this impulse current on the switch (IGBT) and input DC source during starting and normal conditions? how far this is a problem for the SCMLI.
Thank you for your valuable comments
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hi,
A multilevel inverter is a power electronic device that is used for high voltage and high power application because of its characteristics of generating a sinusoidal voltage based on several DC voltage levels.
A switched capacitor (SC) is an electronic circuit element implementing a filter. It works by moving charges into and out of capacitors when switches are opened and closed. Usually, non-overlapping signals are used to control the switches, so that not all switches are closed simultaneously.
for more info:
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Real time PCR related question
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Yes. The dye is made for qrtPCR. According to Promega, the dye has very similar spectral properties to those of SYBR Green (https://www.promega.com/~/media/files/products%20and%20services/islides/is022-gotaq_real-time_pcr.ashx). Any instrument capable of extiting and measuring Fluorescein and/or SYBR Green should work with BRYT Green as well. To my knowledge, this applies to all available qrtPCR instruments on the market.
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We know that robot pose errors are very common and that robot trajectories are pre-constructed and generated from CAD models in machining scenarios. However, for the trajectory points, there are inherent errors, and we need to compensate for the position and attitude of these trajectory points.
The existing off-line compensation is very common, but it lacks real-time, and the compensation objects are all the results obtained from pre-experiments. In fact such compensation, in a new experiment, the whole process is different from that of the pre-experiment because of the compensation done, so the final compensation also all stay at the level of being able to improve the performance, and theoretically such compensation is also all incomplete.
How to compensate for the new point errors based on the information obtained in real time, and update the point information of the generated trajectory in real time?
My robot is an ABB, and it would be great if you could offer some advice on the robot control system,transmission of data, branching of the perception model, etc.
Thanks to all the researchers who discussed and replied.
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Hi everyone ,I am working on three autophagy genes I want to calculate the efficacy so I dilute my samples in this way:1/10,1/100,1/1000 and so on.But the dilutions more than 1/1000 have primer dimers in melt curve,Is there anyway to calculate the efficacy of primers?!
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Please anyone can help me with primer dimer in standard cuvre?!
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Hi researchers
I am doing real time for some autophagy genes .
At first some of them didn‘t have any bonds or had primer dimer!! So I have reduced primer concentration ,increased the template concentration,changed annealing temperature and so on…
Now ,however most of them are corrected but some of them have primer dimers yet, even more than the real bond !!!
what should I do now?!Does any one have an idea?!
I am really confused
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A universal primer annealing feature reduces optimization steps and allows for co-cycling of different assays. A unique combination of innovative buffer, high-performance Taq DNA polymerase, and superior hot-start technology enables exceptional PCR results.
1. avoid universal primers.
2. Host DNA polymerase master mix.
3. Reduce primers.
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Hi,
I have two groups of treated and untreated samples. The biological repeat is two, so I have two groups with a sample size = 2 of each group. I calculated the dc, ddc, and 2^-ddc of treated and untreated groups. I would like to know if the gene of interest is significantly up/down-regulated in the treated group.
Now, I need to run a statistical test to determine the significance between treated and untreated samples. I know that the sample size is too small, but I could not help it!
Since the sample size is too small, I consider using the Mann-Whitney and Kolmogorov Smirnov tests as nonparametric tests. Besides, I have tried the unpaired t-test with Gaussian distribution.
I have tested dCt and 2^-ddCt.
Now, I'd like to know which test you suggest using and what kind of data (dCt, ddCt, and 2^ddCt) is better to test.
Thank you!
Sara
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Thank you for your reply. It's still not clear to me how this applies to quantitative variables, like for instance gene expression measurements. A corresponding variable in bahavioral science might be something like "time spend at some place" or "distance moved (upon some stimulus)" etc. How can ony analyze such data using contingency tables?
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I want to know why we dilute the cDNA for Real time expression studies of microRNA expression.
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For miRNAs from cells or tissues, the cDNA is routinely diluted 1:5 to 1:10 depending on the initial RNA concentration to ensure a final cDNA amount around 10-100 ng per reaction
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When we isolate the edna from water sample generally 2-5 ltr water filtered and dna is extracted but in my case the concentration of dna is very low (7-8ng/ul) and there fore I am having problem in the amplification in pcr. So what is the minimum quantity required to check the amplification using qPCR mechine?
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Thank you @Zulaykha Khurshid for the replying
But the problem I am facing is that I can amplify my gene of interest with semi quantitative pcr with 20 ng/ul but if I use this concentration in real time the gene of interest shows undetermined
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Seeks for analysis reasons to use more recent datasets possible and why not in real time ?
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Dear Abdelmadjid,
can you clarify what kind of data you need?
It is important because there are many repository of real-time medical data however it is not known that you need this kind of data.
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Generally radar detection algorithms to work in real time would require less computational complexity. CA-CFAR is good candidate algorithm in the presence of noise and when electronic counter measures/intentional interference is present, multiple detection comes in the R-D map. We have collected some real time data from mmWave sensor in the presence of ECM. Can any one of you suggest, which one is a good detection algorithms to detect the target in this scenario?. Thank you very much.
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Track the jammer, and use information from that track to reduce the effects of the jammer on other tracks.
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Hello Researchers...!
I was wondering about the most important features in a real-time IoT dataset for machine learning models? any suggestions are welcome and appreciated..!
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Seven necessary properties of highly secure, network-connected devices: a hardware-based root of trust, a small trusted computing base, defense in depth, compartmentalization, certificate-based authentication, security renewal, and failure reporting. https://www.microsoft.com/en-us/research/wp-content/uploads/2017/03/SevenPropertiesofHighlySecureDevices.pdf
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  • I am using the ingredients of 20mM Tris HCl, 10mM (NH4)2SO4, 50 mM KCl, 8mM MgSO4, 0,1 % Tween, 0.8 M Betaine, 1.4 mM dNTPs mix, FIP/BIP primers 1.6 µM, F3/B3 0.2 µM, Loop F/Loop B 0.4 µM, Bst polymerase 8 U, dye(50X) 0.5 µl. Everything was properly mixed and kept in thermal cycler with set temperature of 65 0 C for nearly 100 cycle. I could not able to see the amplification phase, most of the time it follows flat curve and some times with noisy. I am unsure on the reaction part, really it happened or not.
  • The primers works perfectly with the commercial kit, when I implement with real mix of ingredients is the problematic.
  • So please share your knowledge to solve this problem in LAMP analysis on real time detection. any suggestions on components change, concentration change, thermal cycle change?
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Lavanya Malini, Yeah primers is not a problem here. I will try with new batch of chemicals.
Thank You.
Nivedhitha
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I am doing research and as part of this need to develop model real-time model
for sediment study but not getting how to start
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Start with a simple question. First problem: reliable data about sediment, collected on a consistent basis, is hard to find. Begin by writing a simple questionnaire based on what you already know about sediment. Ask yourself a simple question such as: 'What was the proportion of biological input to this rock sample, ranging fro very high for a limestone to very low for an evaporite, with shale somewhere in the middle.' Then collect 1000 rock samples at random from your target area. Now, list your personal 'quick look' percentage estimate of its biological content for each numbered sample . Next ask 100 colleagues and students each to make the same quick look estimate of biological input for each of 10 samples, with their numbers chosen at random by each anonymous participant; note and list each percentage estimate, until you have 1000 numbered estimates. Finally, write a simple program to analyse your 2000 data points using MS Excel.
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How can we map or analyse the real time flood inundation mapping?
Many hydrodynamic modelling needs validation . However, due to lack of data, it could not be more accurate/precise. If we could know the actual/real time flood map, it would be overwhelming and efficacious approach to validate the model which can be helpful for decision makers and disaster authorities.
Thanks.
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Use Google Earth Engine platform
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Which one technique is more applicable for real-time measurement of semantic similarity and semantic clustering? For example, classifying students' answers during the online session into different clusters based on their similarities. Sentences could be from any domain.
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I think for sentence clustering it is better to use Sentence-BERT model a modified version of BERT based on siamese network, which very suitable for sentence clustering. Please refer to the original paper of sentence- bert
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We want to measure real-time complex dielectric properties of LTCC ceramic pellets at temperatures above 500 degree Celsius. Is it possible to measure the complex dielectric permittivitty of ceramic pellets at elevated temperature above 500 degree Celsius using an LCR meter?
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yes you can
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Does MATLAB have the capability to connect any type of data acquisition system with Real-time controller?
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Dear Awad
of course! Matlab/Simulink environment has different functions for real-time applications. I did this for PLC Siemens S7 and Arduino as I/O platforms. If you have any questions, please do not hesitate to contact me with this e-mail: sa_rafatnia@sut.ac.ir
Sincerely, Sadra
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To count objects in real time, I read that YOLO was usually used. In this blog (https://blog.netcetera.com/object-detection-and-tracking-in-2020-f10fb6ff9af3), it says that SSD is an alternative but there is also somes techniques being used that are too slow for tracking in real time in this blog. Is there anything else ? Can't CNN be used in real time for now ?
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Yeah, it is also a good option, Sir Bruno Martin
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Please suggest some research work that states how using just one or the minimum number of sensors to detect which faucet (tap/shower/toilet) is being used in real-time based on water consumption pattern.
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Hello everyone,
Greetings!
Hope everyone is fine and in good health in the current Covid19 pandemic.I would like to know the difference between the Reverse Transcriptase-qPCR and normal realtime qPCR.If we can extract DNA directly from the cells and then prepare the samples for qPCR then why do we go for Reverse Transcription step to prepare cDNA and then qPCR?Kindly explain in detail.
Thank you,
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Hello Abdul Rouf
RT-PCR, also known as Reverse Transcriptase PCR, typically measures RNA expression levels. In RT-PCR, complementary DNA (cDNA) is made by reverse transcribing of the RNA template with the enzyme reverse transcriptase.
This technique is used to qualitatively study gene expression and can be combined with real time PCR (qPCR) to quantify RNA levels. RT-PCR is used in research laboratories to study gene expression, for example in experiments to distinguish exons from introns, and can be used clinically to diagnose genetic diseases and monitor drug therapy.
Also, RT-PCR is commonly used in the diagnosis and quantification of RNA virus infections (e.g., human immunodeficiency virus, hepatitis C virus, corona virus) and the analysis of mRNA transcripts such as those produced by translocations commonly associated with non-Hodgkin's lymphomas, leukemias, and sarcomas.
Gene expression profiling is likely to have a major impact on molecular diagnostics and will depend on RNA analysis using RT-PCR and possibly high-density arrays.
So, it is not always extracting DNA directly from the cells and then preparing the samples for qPCR. Sometimes, we have to go for reverse transcription step to prepare cDNA and then do qPCR.
Hope this information helps!
Best Wishes.
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Hi,
I am working on real-time video captioning using deep neural network, and I want to buy a laptop (not desktop) to execute my work and I need to know the minimum hardware specifications required because the pc with GPU is very expensive as the GPU is the key point.
For example, are the following specifications sufficient or do I need more?
core i7 / 8th gen.
RAM 16 GB
SSD hard 256 GB
GPU GTX-1060 6GB RAM (key point)
thanks
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Thank you
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1. Considering Two Area Four Machines system, I would like to get some PMU data from Area 1 to Area 2. I am interested in finding out the maximum communication delay that can occur so that inter-area oscillations can be damped out by a controller.
2. If controller receives the data after very long time due to channel imperfections, then what is the possible action to be taken to damp out inter-area oscillations?
3. Does anyone has the knowledge on the practical value of communication delay in real-time power system(smart grid)?
Thanks in advance!!
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I suppose peak cancellation CFR(PC-CFR) is now a feasible method concerning to hardware implementation. But when evaluating TM2.0A example of matlab 2020b 5g toolbox in which there is a fast freqency content switching of waveform in each slot(shown in figure), I suppose it's hard to know the frequency content of the signal in advance or recompute the new cancellation pulse coefficients in real time since each set of cancellation pulse coefficients is specified for certain carrier configuration of waveform. Is there any good solution to solve the problem? Or is there any other CFR algorithm which is feasible in this application secene with not much difficulty in hardware implementation?
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probably it will increase delay
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We have isolated linalool and examine by UV , But UV is not satisfactory. if any body is working in the same field i need the real time spectra .....you cooperation is highly appreciated.
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How to develop a suitable model based on problem. Give some real time example
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I agree with @Imra. Nevertheless I throw in a couple of examples. See the attached. Best wishes David Booth see the following response
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How long after radiation of microRNA transfected cells I can do real time PCR test?
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Transiently transfected cells are usually used for Real-Time PCR 24-96 hours after transfection.
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Does anyone know if I can connect the attached DAQ (CASSY type) to MATLAB real time simulink for controlling purposes?
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It depends on your sample frequency, which is limited for performance of DAQ. another issue should be concerned is the number analog of a processing control system. the quality of data will be bad or real-time data might be good if you collect as much data in process as possible.
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hi everyone,
Where can i get free access for real-time historical stock data, or intra-day stock data, which is stock data with every second transaction .
Thank you.
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Dear Rosita Husain,
Greetings!
You can check the following links
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Hello everyone,
I am trying to obtain actual distance (in meters) using the 3D point cloud. I cannot consider directly the z-axis value as the point cloud is obtained from monocular camera.
It would be great if I can get some ideas on how to proceed?
Best regards!
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Hello, I would like to know your opinion about it: apart from checking the amplification curve, what parameters are considered to validate the primers in a singleplex type real-time RT-PCR?
I designed three primer pairs against dengue virus serotypes 1,2 and 3 and plan to adapt it to a CDC real-time RT-PCR assay.
I am new in this type of studies for validation.
Please, if you know, you can send an email: joe_mar02@hotmail.com
Thank you very much.
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Hello Joel,
You can do a melt curve analysis and make sure the primers are specifically amplifying your gene of interest as well as has a single peak. Additionally, you can run an agarose gel for the products and confirm the amplification.
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kindly, provide link if any
thank you.
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Respected Sir,
Refer this link:
Hope this information is useful.
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Which tools are used to extract useful features from network data? The extracted features will be used as input to machine learning models to detect intrusions in (near) real-time.
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Snort. Snort is a very important tool since it is the de-facto standard for intrusion detection systems. This Linux application is simple to set up and can be configured to monitor your network traffic for intrusion attempts, log them, and perform a predefined action if one is discovered.
The most of intrusion prevention systems employ one of three detection techniques: signature-based, statistical anomaly-based, or stateful protocol analysis.
Kind Regards
Qamar Ul Islam
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I developed automatic tuner using Bayesian Optimization.
The plant system is real vehicle.
Furthermore, there is no stability problem, because this system is a kind of semi-active control system.
Is there anyone could suggest a better AI algorithm to be used in real-time calibration of this semi-active control problem with the parameters over 500EA with the real-time vehicle experiments?
This doesn't seems to be a simple problem and needs lots of domain knowledge regarding tuning process and control algorithms. However I would like to find and build a automated tuner imitating what human test drivers do.
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Dear Youngil Sohn,
As you have said so well, this is not a simple problem, for that it will be necessary to choose a tool or a combination of tools which is at the same time powerful and provided with smoothness and this to reach an adjustment including these two criteria. antagonists.
We can offer basic AI algorithms such as Traveling Salesman Problem (TSP), Deep Neural Networks or techniques that are part of Machine Learning.
For more deatils and information about this subject i suggest you to see links on topic.
Best regards
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When I altered the sample time from 5e-7 to 10e-6 in Simulink, my waveforms became severely distorted (solver ode3 and fixed step time in both cases). For real-time simulation with OPAL-RT, I must select a sample time greater than 10e-6. Could someone please assist me with this regard?
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Dear Amit Kumar ,
The minimum sampling rate fs is limited by the Nyquist sampling rate. The sampling Nyquist rate is greater equal 2 fmax where fmax is the maximum frequency contained in the original signal. If the an waveform is sampled with fs=> Nyquist sampling rate you can restore the original waveform by an interpolation process.
So, you have to tell us what is thye maximum frequency contained in your original signal.
If your sampling time is 5 x10_7 s and then you resample it at 100x10^-7 s then you decreased the sampling frequency by 20 times. This will scale down the frequency of the waveform by 2o times. Sure it will lead to great distortion of the signal if it is under sampled.
I do not know how did you decide on such new sampling frequency.
Under sampling leads to aliasing distortion of the signal.
Best wishes
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So here is my problem to solve: People arrive at the airport, first they all go through ID check as one queue. After that, they go through personal security check, and people will choose whichever queue is the shortest at the moment. The personal security check time is unif(0.5min, 1min) and the goal is to control the waiting time within a certain length. Now I need to try different configurations of the number of the people for the security check in order to satisfy the goal of controlling the waiting time. How would I be able to do that?
I now have an arena model that has a decide module in it, set up in N-way by condition, but I have no idea what condition I should give to the module. How would I get the real-time queue length so that the person can decide to go to the shortest one? Thank you!
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Hi, I think you should use the decide function to select N-way by condition and write expressions like NQ(Registration.Queue)<=NQ(Registration2.Queue), the NQ() means the number of people in the queue, and the (Registration.Queue) means the queue of the registration service desk. Hope this will be helpful.
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Hi all,
could you please help choosing the best RT simulator for electrical engineering research.
We are confused between the OPAL (op4510) or RTDS ?
it should have the ability to simulate a complete electrical power system (Microgrids) including generators, transmission lines, PV, wind turbines, relays, arrestors, batteries, super capacitors,....
besides, it will be used in real time control for different electric motors
the easier the programming the best.
thanks in advance
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There are currently many simulators in electronics that are very good. But each simulator is to some extent aimed at a specific type of electronic device. There are also more universal simulators, but in most cases they are for amateur purposes. I think that in order to get good advice from colleagues here, you need to specify for what type of electronic devices you will use it.
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Hi everyone, so I have been running some assays in qpcr(presence/absence experiments).
My experiment is on detection of human malaria parasites.
My positive controls show as present and usually above the target threshold.
However my samples are usually below the target threshold although they yield some curves.
The positive controls have Ct's around 20 and the samples Ct's range from 26 to 30. I use 5ul of DNA for this assay.
I ALSO USE THE ABI 7500 FAST REAL TIME MACHINE.
How do I fix this?
PLEASE NOTE: I have attached some pictures of the results
Thanks.
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Irene-Love Amoah Your samples have curves looking like obtained with significant inhibition of PCR. You can try two-three different protocols/kits for DNA extraction.
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I want to do some research on the application layer in VANET. Precisely, I need the real-time location(the information of gps) about the car. I also need the car and RSU have some power to run algorithm. Which simulation software can help me to finish the experiment?
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veins is ok. you can ns3 simulator aswell
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I am interested in studying the gene expression using Real time PCR and looking for a method that works for isolating RNA from mycobacterium marinum biofilms.
Kindly share the protocols
Thanks in advance!
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I recommended CTAB with mycobacterium it works good.
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The goal of the academic research unit is to connect similar research ideas with technical expertise and funding to facilitate the generation of well-designed, ethical research. Historically, government and philanthropic organizations have been the main sources of funding driving research efforts at academic institutions that enable the generation of fair and unbiased scientific knowledge to the benefit of all its citizens. Institutional oversight ensures strong research- and ethical policy adherence within the borders of its mandate before any research is undertaken or subsequent findings published in formal, peer-reviewed journals. To the fullest extent, this process has resulted in the steady, albeit slow growth of a large, credible scientific knowledge base. Alarmingly, it has become evident that the era of "Big data" has overturned this important scientific paradigm.
The explosion of social media, the proliferation of digital computing devices, and almost universal internet access have resulted in the generation of massive amounts of data that in practice can be analysed by anyone at near real-time speed without institutional or governmental oversight, creating a slew of new ethical challenges. Proprietary algorithms developed by corporate incentives process massive amounts of data to reveal information about emerging social, economic and health trends that inform market strategies and product development. Currently, it is not known to which extent this information is shared or by whom.
These technological advancements offer both opportunity as well as challenges to academic researchers. Given the speed and the amount of data being generated today, are we as academic researchers able to keep up with industry-generated knowledge and standards while bound to the confines and processes of our oversight committees and current funding models? What are the obstacles and opportunities facing the academic research milieu that ensure its relevance in the age of technological advancement? This conversation has just started. Please share your thoughts and ideas below.
Do you agree? If not, can you provide better insight?
In the end, it will be our ability to adapt to our changing environments that will ensure the relevance of the academic research institution over the years to come.
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Certainly not, the traditional style is boring and is not creative. We need a style that is liberated from the routine steps of thinking to reach the conclusion. I don't have the alternative, but someone else may have.
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Between Real time PCR and Nested PCR, which one is the best for enteroviruses detection in stool sample?
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Conventional PCR (C-PCR) has been used to detect specific target genes in various microorganisms (5, 6, 13). Nested PCR (N-PCR) was developed to improve sensitivity but can give erroneous positive results due to DNA contamination @ Buhari Suraka
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Hi, I am currently working on a project which requires me to implement the ICA algorithm in real time, to be specific, I am trying to separate the noise from the audio. When I perform the algorithm in offline, it works fine despite the amplitude of the separated audio is a bit soft. However, when I implement it in real-time, the separated audio becomes very soft. Any source code that I could refer to solve this kind of problem. Thanks.
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What you are describing appear to be two different problems, one compounding the other
For the first problem
1) Which ICA implementation are you using?
For the second problem
1) For the real time capture what sampling rate are you doing the capture?
2) what are you using to capture the signal in real time(e.g. are you using ALSA on Linux or are you using Windows)?
Note that the RM Cortex-M7 of the Teensy 4.0 is a slow processor compared to an Intel or AMD
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I basically haven't found any relevant research, but I think if it is data from surgery or implantable robots, is there a requirement for real-time inference/computing? I haven't found any evidence to support my thoughts
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Thanks a lot for gracious participation respected Dr ranjeet
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I'm doing my Master thesis on improving self-navigation for customers in supermarkets using RTLS (make it easier for them to reach their expected products) then how to apply data collected from RTLS into Machine Learning and AI to improve customers' experience. I'm wondering which type of positioning techniques should be used, Wifi, UWB, BLE etc.
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Phuong Bui Attached document is about AI , Wifi and IoT. may be relevant for indoor positionning.
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In RSA cryptosystem, we generally take 1024 bits long prime numbers p and q. Is any problem if we take 512 bits long prime numbers? What are the security issues may be generated in real time scenario?
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I need codes to encrypt and decode images in Matlab for the following algorithms.
AES, with key 128 bit
AES, with key 256 bit
RSA, with key 1024 bit
RSA, with key 2048 bit