Is it possible for the MRNA covid vaccines to be reverse transcribed? Do humans naturally have reverse transcriptase or do they only come from retroviruses? Thank you!
Hello, Thanks for your question. Can you please reach out to our technical support team at thermofisher.com/askaquestion. They would be the best team to assist. Thank you!
Hi Shekhar! Thank you for your question. We do have a white paper that discusses some of the inhibitors of reverse transcriptase. These include alcohols, salts, detergents, and heparin. Our SSIV enzyme in particular has been tested against these inhibitors and is more inhibitor tolerant than other reverse transcriptases. I have linked to it below for your reference. tools.thermofisher.com/content/sfs/brochures/superscript-iv-reverse-transcriptase-white-paper.pdf
Hi Ale. Thanks for your question. Reverse Transcriptase is not able to add nucleotides without some type of primer. An RT primer is required for reverse transcriptase to build upon and extend. Without it, reverse transcription is not able to occur. If you have any additional questions, please reach out to our technical support team at thermofisher.com/askaquestion. Thank you.
Thank you for your question. Our genomic DNA is transcribed first into mRNA. During the processing, the mRNA is then spliced and the unnecessary regions are removed. This mature mRNA is then reverse transcribed into cDNA. If we were to start from genomic DNA, then we would have those additional regions. Depending on what the downstream process is, this may be fine, but if you are to study the levels of gene expression, one should start out with mRNA, rather than genomic DNA.
Hi Saso! Great question. cDNA synthesis occurs in a 5’ to 3’ manner. The reverse transcriptase enzyme begins on the 3’ end of the mRNA, but is always adding nucleotides from 5’ to 3’ on the new strand. It may look like it’s going 3’ to 5’, but that’s when you’re looking at it from the mRNA strand. The new strand is always synthesized from 5’ to 3’. For any additional technical questions, please feel free to contact us at thermofisher.com/askaquestion. Thank you!”
@@thermofisher Thank you so much for the answer. I see my egocentric thinking in regards to RNA. If I may I have a follow-up. When reverse transcriptase synthesizes cDNA. I am not sure if an enzyme would replace uracil with thymine or would RNA be discarded and DNA polymerase would create "ccDNA" to cDNA. I feel the latter is more logical. Thank you.
Thank you for your additional question. The reverse transcriptase enzyme adds A/G/T/Cs to the cDNA being synthesized. It does not replace uracils as cDNA does not have any uracils present. If a uracil is present on the RNA strand, then the RT enzyme will add a adenine to the cDNA strand. If a thymine is present on the RNA strand, then the RT enzyme will add an adenine on the cDNA strand. I hope that makes sense.
@@thermofisher but there is no thymine on the RNA strand, it's always uracil right? I think they are asking about first single-strand DNA synth where it is an RNA-DNA hybrid from RT, followed by the dna-dna polymerase of qPCR
Really like how examples of Reverse transciptase, Primers, etc are also mentioned here!
wwwooowowo ok so i know what none these things are so im gonna go get some knowledge and come back
This is probably not the place to say this, but you are beautiful!
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I understood thanks for description in very simple words 🤗🤗
We're glad it was helpful! Thanks for checking out our video.
Lovely music
fantastic video - really helped explain things well. Thank you
You're welcome, Ifrah! Thank you for watching!
thank you guys!
Thanks for watching.
thankyou so much your explaination is so easy to understand
You're welcome! Happy to be of help.
wonderful! thanks
A like for helping me find info in my dissertation
Thanks a lot.. Very informative video.
excelente vídeo, entendí todo!
instaBlaster
Super wonderful video THANKS
Thanks for watching!
Thanks!
Good , its help me to understand my task
hey, how exactly do you design gene specific primer for RT?
Is it possible for the MRNA covid vaccines to be reverse transcribed? Do humans naturally have reverse transcriptase or do they only come from retroviruses? Thank you!
“We also show that BNT162b2 mRNA is reverse transcribed intracellularly into DNA in as fast as 6 h upon BNT162b2 exposure. “
I linked the study but UA-cam seems to censor it. Or maybe alex berenson is a UA-cam censor
Just read study saying they do! Like the person above said ^^
Look into what functions the liver can perform (perhaps not the best wording, but the keyword is 'liver').
Thanks dawg
No problem!
great video, thanks!
Thanks! We appreciate your comment.
Hie Thermo, l wanted to know which Qubit kit can l use to quantify cDNA post the Reverse Transcriptase PCR step?
Hi Ian, You can use the Qubit dsDNA Kit. There is an High Sensitivity (HS) and a Broad Range (BR). www.thermofisher.com/order/catalog/product/Q32854
Thanks
Can I use the same primers for both RT PCR and conventional PCR?
Hello, Thanks for your question. Can you please reach out to our technical support team at thermofisher.com/askaquestion. They would be the best team to assist. Thank you!
I have one more question that what is the inhibitors of RT?
Hi Shekhar! Thank you for your question.
We do have a white paper that discusses some of the inhibitors of reverse transcriptase. These include alcohols, salts, detergents, and heparin. Our SSIV enzyme in particular has been tested against these inhibitors and is more inhibitor tolerant than other reverse transcriptases.
I have linked to it below for your reference.
tools.thermofisher.com/content/sfs/brochures/superscript-iv-reverse-transcriptase-white-paper.pdf
Hey i have a question about the primers, what is the purpose of the primers on reverse transcription?
Hello, I searched that up and as I thought, reverse transcriptase requires a DNA primer to start
Hi Ale. Thanks for your question.
Reverse Transcriptase is not able to add nucleotides without some type of primer.
An RT primer is required for reverse transcriptase to build upon and extend. Without it, reverse transcription is not able to occur.
If you have any additional questions, please reach out to our technical support team at thermofisher.com/askaquestion. Thank you.
why use rna first and not directly the dna ?
Thank you for your question.
Our genomic DNA is transcribed first into mRNA. During the processing, the mRNA is then spliced and the unnecessary regions are removed. This mature mRNA is then reverse transcribed into cDNA. If we were to start from genomic DNA, then we would have those additional regions.
Depending on what the downstream process is, this may be fine, but if you are to study the levels of gene expression, one should start out with mRNA, rather than genomic DNA.
Additionally, this protocol is used for research in virology, with so many viruses out there who has RNA genome
Hi. I was wondering why does reverse transcription go from 3' to 5'. Instead of usual 5' to 3'.
Hi Saso! Great question.
cDNA synthesis occurs in a 5’ to 3’ manner. The reverse transcriptase enzyme begins on the 3’ end of the mRNA, but is always adding nucleotides from 5’ to 3’ on the new strand.
It may look like it’s going 3’ to 5’, but that’s when you’re looking at it from the mRNA strand.
The new strand is always synthesized from 5’ to 3’.
For any additional technical questions, please feel free to contact us at thermofisher.com/askaquestion. Thank you!”
@@thermofisher Thank you so much for the answer. I see my egocentric thinking in regards to RNA.
If I may I have a follow-up. When reverse transcriptase synthesizes cDNA. I am not sure if an enzyme would replace uracil with thymine or would RNA be discarded and DNA polymerase would create "ccDNA" to cDNA. I feel the latter is more logical. Thank you.
Thank you for your additional question.
The reverse transcriptase enzyme adds A/G/T/Cs to the cDNA being synthesized. It does not replace uracils as cDNA does not have any uracils present. If a uracil is present on the RNA strand, then the RT enzyme will add a adenine to the cDNA strand.
If a thymine is present on the RNA strand, then the RT enzyme will add an adenine on the cDNA strand. I hope that makes sense.
@@thermofisher but there is no thymine on the RNA strand, it's always uracil right? I think they are asking about first single-strand DNA synth where it is an RNA-DNA hybrid from RT, followed by the dna-dna polymerase of qPCR
ChoOn!!
Perez Maria Thompson David Moore Richard
wrong subtitles and should be RNA not mRNA on the slides
it's true. ın the rxn we usually use mRNA because of modifications. Poly A makes the molecule specific. poly T binds to poly A and starts the rxn
No it's true because reverse transcription is the process of making cDNA from mRNA
100 % lost !!
Conspiracy theorist lol
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