What is RT-PCR? (Real-Time PCR & Reverse Transcription PCR)
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- Опубликовано: 30 июн 2024
- RT-PCR can mean either Real-Time PCR or Reverse Transcription PCR. In this video, I explain how both techniques work, and what kinds of questions can be answered with each technique. This video is an introduction to the conceptual basis of both RT-PCR techniques, it is not a practical how-to.
0:00 Introduction & You need to know how PCR works
0:25 Disambiguating RT-PCR: Reverse Transcription & Real-Time PCR
0:58 What is Reverse Transcription PCR?
2:20 What is Real-Time PCR?
5:18 Results from COVID-19 study that used RT-qPCR
The paper I discuss in this video is open access and you can find it here: S. Deinhardt-Emmer, et al., Early postmortem mapping of SARS-CoV-2 RNA in patients with COVID-19 and the correlation with tissue damage. eLife 10, e60361 (2021). doi.org/10.7554/eLife.60361
Script review by Sheeva Azma (Twitter @SheevaAzma)
Photo of Terri Sewell getting COVID-19 test by the United States Congress, Office of Terri Sewell - Public Domain
Video of using Real-Time PCR machine and pipetting into 384-well plate by Allie McClure and Berta Canal, used with permission.
This video is copyright Jayme Dyer, published under the Creative Commons CC-BY SA 4.0 license.
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i dont know why i am studying at a university WHEN THESE CHANNEL ARE HERE ON RUclips . really ggood
I totally agree
I'm suspecting this will be one of those channels that will be incredibly successful in years to come. Growth will be slow especially on RUclips, but if you continue producing incredibly informative and quality explanations like this, I'm sure you will go far. You explained this concept perfectly. Thank you!
Thank you for the lovely compliment! I'm glad my video was helpful.
The MIQE guidelines published in Clinical Chemistry in 2009 clarify this issue: RT-PCR is reverse transcription PCR. Real-time quantitative PCR is qPCR.
Being new to PCR and stuff, I was really finding it confused , especially after going through the literature. But here? It's a wow, never I had come across of such an explanation which not just cleared my confusions but paved a way for some novel research ideas. Ton of thanks. Warm appreciations and looking for more of this kind. Thanks again. Good day.
Thank you so much for this!! I needed to understand how exactly real time pcr worked for a case study I have to present in front of pathologists for school.
Incredibly clear, engaging, and informative video! I needed a quick refresher and I now am totally comfortable with the concept of RT-PCR and qPCR. Thank you for making this!
Thanks! 🥰
OMG.
Loving your explanation (and thanks for it), the music, and the dynamic of the video.
Thank you so much! You are the best explainer ever. Plus, you have such great energy and demeanor, you are also fun and pleasant to watch and to listen to. I hope you are a teacher!
Thanks! 😊 I am a teacher!
I love how easy this makes it to understand
great video! its a talent to explain so complicate things, so easy! Congrats!
The best exaplanation so far
Everything here is lovely explained, thanks!
This was so helpful. Highly recommend!
Thank you very much for the simple and concise explanation
Extremely thankful.
Wonderful explanation.
Thank you for such an informative and educational video! This helped me learn the difference in RT-PCR and PCR for my Virology exam tomorrow!
Great! Glad it was helpful!
You made it crystal clear mam ..thanks a lot . please make more explanatory vdos
Thank you for the explanation. They are really clear
Just WOW !
Excellent explanation really !!
Thank you❤️❤️
Thank you!! 🥰
Amazing video! Keep up the great work.
Thank you so much for this!!
This is super helpful. Thank you so so much for doing this
Glad it was helpful!
this was excellent. thank you.
Wonderful explanation
Great video, easy to understand! thanks
Glad to hear!
good use of real life examples
great vid thank u!!
AMAZING THANK U SO MUCH!
Amazing content!
Amazing example of scientific communication!! Thank you from a fellow science communicator.
Always special to get a compliment from another science communicator!
thank you so much!
🎉🎉You are good a teacher
Very profitable video thank you ❤
Please make more such amazing videos.
Thanks! Will do
underrated video
Thanks ❤
awesome
Cheers Jamie :D
I owe you :( super thank you
Do you have a video explaining amplification techniques such as FRET, TaqMan, Molecular Beacon, and Scorpion Probe? HELP!
Sorry if I sound incorrect here, but through explaining Reverse Transcription PCR you explain how when you have regular DNA at the cDNA stage, does that imply you have left the Uracil in it, with not using RNAse H and Extending the DNA from the fragments to create a double stranded copy DNA of the original mRNA?
the RT PCR Test used for Covid-19 - Though it is easy to trace the discovery of Reverse transcriptases to Howard Temin , and its independently isolation by David Baltimore in 1970, and it is also very easy to identify the inventor of underpinning PCR Test as Kary Mullis, I have yet to identify a scientist or group of them that take the credit for each of the RT PCR Test (real Time vs. Reverse Transcription). Do you know? Does anyone know?
So can you use primers for specific genes that code specific proteins and then use RT PCR in both its meanings to quantify how much there is of each mrna?
i guess so, because that's exactly what i'm doing in my masters degree haha
Helloo, so we know that DNA polymerase cannot use RNA as a template to build a complementary strand. What is the biological reason therefore? Is it due to structure or direction of activity?
You are not showing on your graph the number of copies. Like you said ten versus 10000 is not shown on the graph. I get the guicker plateau in fewer cycles and that of course means more copies for that sample, but it's not plotted on the y axis.
After complete reverse transcriptase and real time pcr,how we test the sample, validating it has sars cov 2?
If there is PCR product after reverse-transcription PCR, then there was SARS-CoV-2 in the original sample. If there is no PCR product, then there was no SARS-CoV-2 in the original sample.
can you recommend for me a book to understand the extraction of alkaloids or extraction in general?
and thanks
Interesting question! Unfortunately alkaloid extraction is outside my area of expertise, so I can't make specific recommendations. :(
@@YouTooBio
Ok.
So, any recommendations about practical books in your field?
@@ahmadhegazy7811 I can't recommend specific books in my field - we tend to learn techniques through direct mentorship from other grad students or postdocs in the lab. However, if you're looking for step-by-step video tutorials, definitely check out Jove www.jove.com/
@@YouTooBio thanks 🖤
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shere material please