Science method
Genetic Engineering - Science method
Directed modification of the gene complement of a living organism by such techniques as altering the DNA, substituting genetic material by means of a virus, transplanting whole nuclei, transplanting cell hybrids, etc.
Questions related to Genetic Engineering
We have been using Bt toxins for a long time, if a situation arises that the targeted organism becomes resistant to that toxin will it be possible to kill the targeted Bt resistant insect with the chemical insecticide that we use to kill the non resistant one. And if not what are the steps we can take? Definitely one of the options will be more toxic insecticides which will result in killing of a broader section of non targeted organisms which will effect in disturbing the equilibirium of the environment. Are there any other suggestions?
Is it necessary to use any kind of Tag ? what alternative methods could be used?
The first transgene will be easily screened by using marker gene analysis, but in case of second, 3rd and 4th genes it may give false results (positive results), then what alternative methods could be used?
I am trying to find a cis element such as a/ttttgxxrttaagg, in this case, is there any program on-line that can search my target sequence and find out the matched cis element? Thank you.
I am researching in construction of recombinant luminescent strain, but know nothing about how to choose a suitable host cell. Any advice?
I want to co-express two different genes in the Pichia pastoris, so as to catalyse a biological conversion from substrate A to intermediate product B, and to the final product C using the two expressed enzymes. I don't know which kind of plasmid can be used, and also want to know the way constructing the recombinant plasmid.
Have there been any identified side-effects of transgenesis? Do you agree ethically with this biotechnology especially in animals?
Point mutation of ssrA gene is very very rare phenomena, cause which is ribosome rescue factor/molecule act as both tRNA and mRNA during error in translation, i done lot of attempt but i cant, I clone ssrA gene into multicopy plasmid and mutagenesis on cloned ssrA gene, now i want to introduce mutant fragment of ssrA in multicopy plasmid into chromosomal region, i cant able to do this, anyone suggest for this?
I want to establish a cell or cell line to secret BMP4 permanently and I haven't any practical experience about this. First I need to know which kind of cell is appropriate for this purpose?
And I think I need appropriate vectors and cDNA that lead the cell to secret BMP4.
Are there protocols out there, that could help me out with that task? Expression should be in human cells under the control of a minimal promoter, assay for promoter activity would ideally be GFP.
I have to clone and express a viral gene in a mammalian cell line so I'm searching for a shuttle vector including eukaryotic elongation factor 1 alpha (eEF1alpha) promoter with multiple cloning sites plus neomycin & ampicillin resistance gene for selection. Does anyone have experience with such a vector?
My gene has been sequenced from genomic DNA of the target species and I already have tried altering Tm, Cycle, Duration of each steps, cDNA+Primer+Mg salt concentration. the reagents are totally ok.
I extracted the RNA with Trizol and RT kit was invitrozen. What could be the problem?
Thanks in advance.
I designed a construct of VP16 activation domain fused with a HD domain of a transcription factor (the transcription factor is on the C terminal fo the VP16) in an expression vector. However, the fusion protein did not seem to initiate transcription activation. Does that mean that VP16 activation domain is not enough, that additional domains of VP16 are required for recruiting other factors for transcription? I'm puzzled with how this system works.
I find it difficult for citizens to get the best or the worst out of GMO's due to the unfavourable discussions I have come across. Help me out somebody, is it good or bad?
Hi, I'm trying to do a 5' RACE but I'm not getting a clear product. It is very smeary and the amplicon is not even the expected size. I am using the "5' RACE System for Rapid Amplification of cDNA Ends, Version 2.0" from Invitrogen. Primers (GSP1 and GSP2) have been tested on genomic DNA and work properly. I suspect that my template cDNA concentration might be too low.
After SNAP purifying the reverse transcribed cDNA, I nanodropped my sample and it showed 5-8ng/ul, which seems very low. Perhaps the transcript I'm trying to amplify is not expressed at high levels? Does anyone know what the template cDNA concentration of a sample before PCR should be in a successful 5' RACE reaction?
Thanks
I am planning to do a site directed mutagenesis in a 1.8 kB gene. I wish to mutate one amino acid at a time to study its role in enzyme catalysis. Can someone suggest good and affordable kits that are currently available?
Have a look at the special Nature Collection on TAL Effector Nucleases:
I need a fungal gene synthesized, but I'm having trouble in choosing between the existing companies. I've been Google searching, and found two companies in the US that I took in consideration: Biomatik and Genscript. Problem is that I haven't found any customer reviews about them... I was wondering if any of you have some past experience with these companies, or if you could suggest a different company which is good (and maybe cheaper as well) for this purpose. Much appreciated!
Hello there,
I am interested in a gene from sheep, CCT8 (chaperonin containing TCP1, subunit 8 (theta). However, I don't know if it was already sequenced, so I am trying to identify and annotate it manually. I blasted a protein sequence from bulls against ovine sequences. A lot of unknown cDNA sequences from ovine came up. Some of them showed 100% of identity. Which sequence should I choose? Which tools can I use instead of clustalW?
Thanks. Appreciate any help.
I have been working with whole-genome expression data using ESCs and embryonal carcinoma cells, but due to the size of the datasets, I sometimes struggle to easily identify particular markers of differentiation towards dermal layers and markers undifferentiated cells. Other than the typical gene annotation tools (DAVID etc), which I find often misses certain potential marker genes or produces false positives, I haven't found any other trustworthy repository of marker genes besides the tedious process of comparing them to previous papers.
Would anybody know a quicker and easier method of identifying marker genes in the data sets?
Natural selection
Does the evolution of beings by natural selection necessarily show that they are intelligent beings?
Update basic genetics conceptions-comprehensions. RNAs are Earthlife's primal ORGANISMS...
Life Is Simpler Than They Tell Us
http://universe-life.com/2011/08/31/origins-in-cells-clusters-intercell-cleanup/
Evolution:
Genes to Genomes to Mono-cellular to Multicellular Organisms;
Direct Sunlight to Metabolic Energy, Too;
Tryptophan to Serotonin to Melatonin to Neural System. (cue for inter-cell cleanup time)
A. Tryptophan to Serotonin to Melatonin
Melatonin is a hormone secreted by the human pineal gland during night-time darkness. It is now marketed in the US as a nutritional supplement. The hormone is an indolamine compound derived from the essential amino acid L-tryptophan, with serotonin as an intermediate precursor.
Tryptophan is one of eight essential amino acids not produced by the body but coming from the diet. The additional fourteen amino acids are produced metabolically.
In the brain, tryptophan converts to serotonin, the neural-transmitter. Tryptophan is the only source for serotonin in the brain. Insufficient L-tryptophan in the diet is a cause of many severe biological malfunctions.
Some serotonin is converted in the pineal gland to melatonin, the hormone involved in inter-cell processes during sleep time.
B. Sunlight to Metabolic Energy
Bio-clocks are products of the innate sun-dictated active-inactive pattern of genes and genomes, parents of Earth’s life. During life genesis and its early evolution direct sunlight was the only source of their usable energy. This situation persisted well into the evolution of the early mono-cellular organisms, and both genes and genomes display, therefore, innate “inactive-sleep” phenomena.
The incorporation of mitochondria with some cells initiated the metabolic bio production of bio usable energy and furnished the evolving mono-cellular organisms with new, additional, flexibly available local energy. This development opened up a variety of courses of evolutions of cultures of mono-cells communities.
C. Individual Mono-Cells to Cooperative Mono-cells Communities, Cultures
As individual independent genes aggregated to cooperative genes communes, organisms, genomes, so individual mono-cells aggregated cooperatively into mono-cellular communities, cultures.
http://universe-life.com/2006/03/22/natural-selection-at-cellular-level-life-is-a-cooperative-affairs/
“Life has always been and still is a fractal affair, repetition of phenomena on ever more complex scale. It cannot be otherwise; it evolves. And surviving-proliferating life has always been a cooperative affair since cooperation is most successful for overall survival/proliferation.”
Cooperation requires all sorts of interactions, including maintenance, protection and foraging for food-energy. Organisms’ interactions are “cultures”. Cultures require “cultural energy”. Melatonin and some proteins are dark-and-light cue signals evolved by the mono-cells communities for timing inter-cells processes when the intra-cells processes are at “sleep-inactive” state. Melatonin is a derivative of serotonin a derivative of tryptophan, and proteins are genes’ tools, energy-dependent metabolism products.
D. Mono-cellular to Multi-cellular Organisms: Mono-Cells Culture to Neural System, to Nerved Multi-Cellular Organisms
Now we can appreciate the fractal nature of life’s evolution. It is ever-continuous ever-enhanced ever-complexing cooperation. Now we can understand why, and grosso modo how, all the organs and processes and signals found in multi-celled organisms have their origins in the mono-cells communities. And this includes the functions of serotonin and melatonin and, yes, the evolution of neural cells and the neural systems with their intricate outer-membrane shapes and functions and with their high energy consumption requirements.
Now, circa four billion years after initial genesis-evolution with direct sun’s energy followed with evolution with also indirect, bio, sun’s energy, some of Earth life, we humans, find ourselves short of energy and in need of exploiting again more, and more directly, our sun’s energy…
Dov Henis (comments from 22nd century)
http://universe-life.com/
http://universe-life.com/2011/07/10/seed-of-human-chimp-genomes-diversity/
I like to study about genetic distribution of plants.
Baculo virus transfection
maybe M. tuberculosis
maybe M. tuberculosis
Assuming that we have a good negative control, and the right melting temperature, what can cause very low fluorescence level of the melting curve?
Which of the plasmid copy number detection protocol can help? Protocols on net ? Can anyone specify any ideal protocol? Please send some recent reviews or papers even in animal system if you have..A good statistical explanation is what I am also looking for..
Does anyone know software or online tools that could help me make predictions of a protein sequence. I would like to test whether it would be a possible transcription factor.
I need a pBS-SK-Sfi cloning vector. It would be a great help if anyone can provide the same to us.
Can someone help me with the oil <fatty acid> producing strain of alagal species? For example, Botryococcus braunii and its fatty acid producing strain and its properties.
I'm currently trying to figure out if anyone knows of any data on the transcription start site of the human U1 promoter in case of the missing initiator box. Does the -50 element still define the location of the start site in this case? Is the purine rule still valid then?
What protocol do you follow to get the best results? Thanks in advance
What would be the best technique for in vitro cell fate mapping, when the duration is about a month? What different protocols or products are available?
Although I searched online, no information was found.
I was trying to purify some ChIP DNA following several histone modification IPs.
After reverse crosslinking the DNA fragments, I added RNaseA to it, incubated for at least 30 minutes, added Proteinase K with Tris and EDTA and incubated at 45C for over 2 hours. This procedure/sequence of steps has worked before and it's the protocol I follow. Following this, I did a phenol:chloroform:Isoamyl alcohol extraction, followed by a chlorofom extraction. Then I proceeded to precipitate the DNA using glycogen (2µL at 5µg/µL) and 2.5 volume of 100% EtOH.
I saw goop precipitating in some of the tubes I was working with. I treated all the tubes exactly the same, so I cannot comprehend why some of the tubes formed this goo precipitate where as the rest did not. After centrifuging, I removed the EtOH supernatant from the tubes and let the pellet dissolve in 0.1X TE. I'm thinking about repeating the RNase A, Proteinase K treatment and the Phenol chlorofom extraction.
My question is, when repeating the process again, should I be adding more glycogen for the DNA to precipitate, more salt, etc.?! How else do you think I can recover my DNA fragments? Will repeating any of the process I listed above affect my yield?!
Thank you so much for your input! :)
I want to transfer the GFP protein gene to my cell lines. I want know the simplest way to do so. Preferably a chemical method.
Every time I was doing allele replacement experiments, I met this problem. It seems that one of my homologous arms didn't work during the two recombination process. Due to it, all I got finally were wild-type ones, none of mutated-type.
The band may be of 100bp. Can anybody suggest the standard protocol for RE Digestion?
I am looking for PCR conditions to amplify this gene.
Dears I want to produce drug in plants (Molecular farming) by expression my gene in lettuce. i have a challenge, it is enzymatic degradation in the stomach. my question is " Are there any amino acid sequences that protect my drug in stomach?" can i attached this peptide to my protein? Best regard
I'm studying the population of soil microorganisms present in the rhizosphere. I used DGGE to investigate whether differences exist between populations of healthy and decayed trees. I've used the Pearson's Correlation to build the similarity matrix and UPGMA to construct the dendrogram.
Now I would like to select some samples to sequence the V6 (for bacteria identification) and ITS II (for fungi identification) region but I do not know how to do it. Or better ... I have no way of justifying why I choose certain samples.
Does anyone have any idea how to select samples for sequencing based on data obtained from DGGE (Denaturing Gradient Gel Electrophoresis)?
Thank you
hey guys. I am willing for a training in rDNA techniques in India. Please get me the updates, if any.
Friends
I srihari, requested you to get latest updates about restriction endonucleases. If you have any body please send me to sriharigunji@gmail.com. please
I have a question related to sequencing analysis. Does anyone can give me an idea for exp. if I found the same species in my sequences is it good or not to do the phylogenetic three ?
One more question is that how could we know about the revolution of gen by using sequencing analysis or another method is you know ?
Hi,i have completed B.Tech in Bio Technology,during my final year project me and my friend found one complete new remedy for diabetes from INSECTS,by animal studies we confirmed.We got 1st prize in an International Project Competition and Exhibition held at Vel tech college on Feb 2011.We PATENT our project in Intellectual Property of India, our C.B.R. No-7180.We want to study MS in abroad,Could anyone help us?
If i find right professor i will be grateful to you people,If anyone knows a professor who is working on diabetes project or going to start diabetes project kindly inform me. We are ready to come any part of the world to study and research. Thanks you people who all are read my post.Take care.
I want to design my Ph.D project on this.
Dear friends,
Im using cell designer for pathway drawing. I finished it. Then i have to analyse the path. For that i need to import the .xml file into cytoscape. I cant. Some error was occuring. anybody had any idea , please help me...
or suggest me some tools for metabolic network analysing...
Thank you.
which is most effective method for seperation of protein and why?can all the protein being seperated by all the method.
talk about biotechnolgy in Colombia is so difficult, because research isn´t as important as talk of poverty, inter conflict, drugs and narcotrafic corruption, because there are the principal troubles in my country, the research in this topic is less avanzed than other countries as Chile, Brazil and Mexico, our action field is limited, there isn´t companies looking like such a biotechnological profile but rather looking for people from other areas to apply biotechnology i am engineer biotechnology now, how is biotechnologies studies or research in Colombia?? this for wind our action field.
can all the target dna combine with all type vector or have specific vector for recombination
my question is that is it possible in the biotechnology to produce a virus with the desired properties .. for example if some one need some receptors or any other molecule or... Show moremy question is that is it possible in the biotechnology to produce a virus with the desired properties .. for example if some one need some receptors or any other molecule or channels or pores which are not normally present in the natural viruses .. answers from the experts will be appreciated...
i want to do my main project in PG.. so please tell me..
hi... im from Malaysia. I really need help from u all. I cant find any journal about dna barcoading for aloe vera. now, i just refer to dna barcoading for land plant. actually.. i m am 3rd year student of chemical engineering in bioprocess and now i need to do final year project. my project is about development of dna barcoading technique in identifying aloe vera.
This relates to drought stress in the tropics.is using Rec. DNA Tech therefore an acclimatization to the high temp stress?e tropics.is using Rec. DNA Tech therefore an acclimatization to the high temp stress?
Journal of Pakistan Medical Students
How does the virus know that it does need to mutate or is it just random? Are there known internal factors within the living organism are the direct cause for mutations?
Why does a substitution in nucleotide base pairs take place?
Or perhaps non living mater that is near by as part of the life support medium of the living organism?
I’ve been asking myself for a long time and perhaps many others still are interested in the Virus mutation mechanisms, more like the factors that produce such event(s). Here, also as branching out the question, I propose to discuss about the biological transmutation of elements phenomenon.
Enzymes suffer transformations, modifications, mutate. Viruses do mutate.
There are mechanisms, the internal or some external clocks and / or some physical events with chemical reactions implications that take place I believe.
Are isotopic activities possible factors to trigger virus mutations? Is it the same for enzymes?
Perhaps we should look at it closer to be able to identify not only possible triggers but also to better understand the mechanisms that mutations take place in their complexity.
Or perhaps we have a combination of isotopic activity and other activities that interact?
If any isotopic activity is involved, what are the isotopes we may talk about or have under observation? What are the atoms we may look closer at?
What is known, proved or at least of hypothetical nature in this area of concern?
I hope I made myself understood as is not really my line of specialty.
Thanks to all members in advance and perhaps we can work something out, a research project may be borne out of this as team work? It is multidisciplinary complex subject that perhaps is worthwhile to try to unveil.
Adrian Toader-Williams
ca anybody suggest me a rare genetic disease to do project work on it....plz provide some information about genetic diseases...I need to do research on it...
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Is there any "Lycopene" gene construct available in public domain ?
Dear friends,
I need the of GFP and cellulose gene constructs, can any one help me in this regard.
Hello, I'm wondering since blackhat hackers abound on the internet, and have plans for even RFID transmitters in national IDs, and break into everything electronic and non (like lock picking) and are experts at social engineering, how nanobiotechnology will deal with the first human/machine interfaces, especially in the area of the brain. I was going to be a whitehat hacker for a while (defense) but it wasn't my vocation and was mostly curious about certain aspects, but basically, a human interacting directly with the Internet or computers, even stand alone, stand the chance of becoming "infected" as well as chips inserted in not just the brain but other areas, when anything from video games to other software have come out with virii on the manufactured DVD/CD, directly from the company. We definitely need safeguards, as well as "firewalls" for our brains and even DNA since that is our code/blueprint for healing, growing and procreating and adapting basically, since our bodies change genetically each day per reading material I've read. Also, spies, terrorists etc could be passed easily down the line genetically if implants are somehow used to change genetics, creating tendencies and even memories. I remember that some dreams/memories or near death experiences that are common are the results of what is in our genes. Even info could be passed on this way similar to hiding code inside a photo, the human body/brain is technically a computer/machine. http://www.tricksystem.com/2008/12/secretly-hide-any-file-inside-jpg-image.html IF someone makes the connection and basically translates the machine code to human code, it's like converting from hexadecimal to base 10 in comparison. Also, stenography is basically how to hide messages and with distributed computing etc and botnets, perhaps people could be controlled or manipulated easier and is dangerous for all, especially military or scientists. Religion already shows that masses can be manipulated (especially when used improperly) as does media and entertainment such as music, movies and even books. The effects of music alone are hypnotic and activating someone, even involuntarily is not impossible nor extremely difficult. Think of something akin to the Manchurian Candidate but without direct involvement per se and no blame except against the individual. Best of luck, am tired and ill and going to rest but I have more ideas and activating individuals, since we're bio-electrical can take as little as radio waves, cell phone microwaves, EMF, an MRI etc. BTW, anyone interested in funding my education? I thought of this with a mere associates in Telecom and 3 semesters until my Telecom. I'll tell you that Ray Kurzweil is one of the people I admire and I've books of his that I checked out and also own "The Age of Spiritual Machines" since he's into nanotech as well as A.I.
Dear friends, This is mohandev from Andhra university visakhapatnam, India and I need a cellulase gene for my work. How can I get that cellulase gene hardcopy and can anyone is provide that consrtruct to me ? please help me i this issue
can anyone please tell me what is the sensitivity (% level) of detection of gm foods by PCR when they are contaminated with Non-gm foods ?
(primers cam 35s, nos, EPSPS )
please mention recent articles possible.
Thank you all !
in fact i look for a linker for hybridization of two distinct gene. how can i find it?
Thanks.
Does anyone know the prevalence of the Mx gene in Egyptian native poultry breeds?
Fayoumi
Does it mean the shape, size and the intensity?
Low weight molecular proteins are easy to be degraded in E. coli :(
We know that the heterochromatin assembly in S.pombe is guided by RNAi, But if we are looking for a gene on euchromatic region i.e. on normal chromosomal locus what would happen?
A question is also raised if we silence a gene of primary metabolism then what would be the fate of cell longevity.
I kept DNA samples (undiluted) which I isolated about 3-4 months back and preserved at -80'c and after 2 months I kept them at -20'c. Yesterday I checked them on gel. Almost all of them show shearing.. I did not save samples also. ..Any suggestions...Do they give amplification on PCR...
This series of articles traces the development of Genetic Engineering from its biological roots through to its biggest applications. The journey starts off with the elucidation of the principles of heredity in the mid-late eighteen hundreds and ends at the cutting edge of genetic engineering today. A. R. Chakravarthy invites you to make that journey.
Knowing the fate is to make preparedness or unwanted worries about likelyhood of onset of particular illness or diseaes.
I have a grant to develop protocol and stably transform our local tomato varieties for disease resistance. I am in search of a suitable laboratory where such work can be done, preferably where bench fee is not charged. The grant covers flight ticket and subsistence.
Who has the vectors, pCL112 and pCL113?
These vector can be used to test protein interaction by BiFC.
It could be Land snails DNA and gene extraction.
Dear all..
I've finished to culture and isolated the pET-15b-56ST-GFP Plasmid... now I'll try to cut the GFP gene from pET-15b-15b-56ST-GFP plasmid, but I don't know with what I supposed to cut it... because there are so many cutting site in there, so I still confuse what the better one of restriction enzym I supposed to used..??
I want to put the GFP gene to pBI121 plasmid as one way easier to predict the gene that we put to the plant is expressing in it... but the matter is I still dont know how to cut the GFP gene, what the restriction enzym that I supposed to used??? can anyone help me to answer it, please!!
Best regard....
if we try to inter GBMs emboli into arterial cerculation(animal models) could they cascade itravascular and adherte to brain vascular ednothetium and make changes in the microenvironment as systematic metastasis and open BBB to survive in the parenchyma causing metastasis
or auto-immunity suppress and kill cells ,although melanomas have the same genitic origin can cross our immunity system and make metastasis,Farther abscess have the keys to open BBB.
thanks.
This is my project title; i was wondering what specific things should i take into account?
So that a lot of info could be found
-thanks-
The banana cv. I work with grows 14ft. in height
Can anyone has a gene for inducing dwarfism in banana, how to get it?
When I run the crude plant extract in SDS-PAGE, I see a clear gel with no protein bands at all... kindly suggest me some ways.
Since hyposia can increased red cell production. I think we can just check if suppression of red cell production under exercise can do this.
If i would like to clone a marine metagenomic library and i would use E.coli as a host for the preparation of metagenomic library then in that case there is chance that i would miss the expression of particular gene function bz it may be possible that it will nt express in E.coli, so how to over cum ths problem????
The land snail is a popular edible mollusk.
Hi everybody,,, any one knows which genes controlling boron absorption and translocation in plant?, specially brassicaceae family
Are some cultivars of wheat with genome of the species T. macha?