Questions related to Real Time
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.
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?
Red: Control enzyme (Hot-start)
Blue, green, purple, and pink: KOD with different buffer.
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
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?
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..
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.
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.
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?
Investments are real-time updated science, due to its special properties. Therefore, tell us what the last updates and topics about it are.
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
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.
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 :)
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?
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.
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.
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.
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?
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?
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?
I am using Step One real-time instrument and the Green master mix with ROX.
Thank you all
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.
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 ?
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?
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?
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
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.
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?!
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
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.
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?
Seeks for analysis reasons to use more recent datasets possible and why not in real time ?
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.
I was wondering about the most important features in a real-time IoT dataset for machine learning models? any suggestions are welcome and appreciated..!
- 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?
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
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.
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.
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?
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 ?
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.
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.
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)
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!!
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?
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?
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: email@example.com
Thank you very much.
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.
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.
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?
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!
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
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
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?
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!
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.
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.
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
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.
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?