Reverse Transcriptase PCR (RT PCR) | What are the applications of RT PCR ? | RT PCR protocol

This video talks about the principle behind RT PCR. Reverse transcription polymerase chain reaction (RT-PCR) is a laboratory technique combining reverse transcription of RNA into DNA (in this context called complementary DNA or cDNA) and amplification of specific DNA targets using polymerase chain reaction (PCR).[1] It is primarily used to measure the amount of a specific RNA. This is achieved by monitoring the amplification reaction using fluorescence, a technique called real-time PCR or quantitative PCR (qPCR). Combined RT-PCR and qPCR are routinely used for analysis of gene expression and quantification of viral RNA in research and clinical settings.
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Thank you so much for these very useful short tips!

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[@davidace7]

very good, detailed and concise explaination, thank you sir!

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@@animatedbiologywitharpan sure sir ... lov your explanation

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Very well explained thanks !! The animations are really good

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@@animatedbiologywitharpan yes I did share it with my friends it is really helpful

[@SuperLinhi]

I have a question: when I have transcribed the mRNA into cDNA, do I perform a normal PCR with the cDNA?
Can someone pls tell me if i'm right or wrong ?
PCR steps
1) denaturation of the template DNA
2) use primer and poymerase
3) amplification
RT-PCR:
1) isolation of RNA
2) transcribe with reverse primer and enzyme reverse transcriptase into cDNA
3) follow PCR steps

[@IronMan-dj1uw]

Hello Bro
You have not included the further steps
Like After making the hybrid rna-dna .The hybrid's rna is partially digest by Rnase H ,then DNA polymerase I forms a complimentary strand and nick is sealed by DNA ligase and
Thus forms a cDNA

[@animatedbiologywitharpan]

Yes these are nitty grity details. It was a overview hence did not get into details

[@mariafernandalealtamez9844]

Very good info! Do you have the references??

[@ikennafranklinokoro4252]

Great video, comparison of RT-PCR and PCR was a benefit

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[@geekymedico.8099]

Amazing explanation ... thank u sir...🙏🙏🙏🤗🤗🤗🤗🤗🤗

[@hidayat.609]

awesome

[@animatedbiologywitharpan]

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[@sangitamandal6146]

It was very helpful ❤thank you sir😊

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I can't thank u enough for this... just keep going 🙏🙏🙏

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[@dhamotharan6745]

Sir can u explain about multiplex pcr

[@animatedbiologywitharpan]

Sure it would be uploaded soon

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Thanks, good content, should give you thumb up.

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[@yokoshiko9427]

Great help on exam eve😇🤩

[@Xahidd____]

Can anybody tell me if reverse transcriptase posses 5'-3' exonuclease activity then why aren't Hexamers degraded instead they are incorporated 😢😢

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very nice channel thanks from turkey arpan

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Please share my channel link with your friends and help me to reach big audience. By the way I love 💘❤ Turkey and the amazing food. I still remember my visit to Istanbul.

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I just love baklava and kebabs

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Thank u sir

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thank you

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Thank you so much sir.

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[@sanjeevyadav1976]

Sir is this technique used for hiv detection

[@animatedbiologywitharpan]

can be used

[@anusreesaha4624]

Please upload a video on allele specific PCR.

[@animatedbiologywitharpan]

Thanks for suggesting

[@sanajiya4462]

Yawrrr such a super duper lecture thank you so much🥹✌️

[@animatedbiologywitharpan]

Pcr playlist ua-cam.com/play/PLKtiwIJ8Q7rpWfwFl7dpbamj09MQ7ynPT.html&si=sCuhziSFd8j-dSPn

[@Yousha999]

Hi thank You for the well explained video can you explain microRNAs

[@animatedbiologywitharpan]

Ya there would be series on all the regulatory RNAs soon

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you saved my todays prentation sir, thank you very much!

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[@ritikyadav3014]

Hello sir, i want to go for research but i don't know how to find research institute for my topic of research can u help me that how can i search institute.

[@prachimate2238]

can u plz explain tma and nasba

[@animatedbiologywitharpan]

Can you tell me the full forms

[@shubhamrana41999]

sir is rt pcr is same technique used in covid rest or it is different .

[@animatedbiologywitharpan]

That RT pcr is real time PCR also qPCR . One step of that is reverse transcription which makes cDNA from RNA . After that qPCR is done from the cDNA.

[@AdityaRaj-zk9eh]

thanks alot

[@animatedbiologywitharpan]

Could you please help me by sharing my contents with your friends group/ college group. I put huge efforts in making these videos but unfortunately not a lot of people are watching this.

[@animatedbiologywitharpan]

Could you please help me by sharing my contents with your friends group/ college group. I put huge efforts in making these videos but unfortunately not a lot of people are watching this.

[@jennymacparry1855]

What if we don't use hexamer primers???

[@animatedbiologywitharpan]

Alternatively you can use oligo dt

[@__jm__6762]

Dont we use a specific primer in RT pcr with those 2 primers?

[@animatedbiologywitharpan]

Generally no. In Reverse transcriptase PCR all mRNA are converted to cDNA . Ideally there should not be any bias. During qRT PCR( quantitative real time PCR ) we use specific primers.

[@__jm__6762]

@@animatedbiologywitharpan thank you so much sir

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@@animatedbiologywitharpan thank you so much sir

[@sabinabegum419]

Inverse pcr k upar ek banayega plz

[@animatedbiologywitharpan]

ua-cam.com/video/vtHwOWMPTL0/v-deo.html

[@myeverydaylife20]

hello dear. The RT-PCR consists in 1 cicle only ??

[@animatedbiologywitharpan]

For reverse transcription protocol yes

[@animatedbiologywitharpan]

For q RT PCR ( quantitative real time PCR ) se the following video ua-cam.com/video/_pMCwREQ22I/v-deo.html

[@myeverydaylife20]

@@animatedbiologywitharpan Thank u so!!!!

[@thisslightlysweetlife3402]

Where does the reverse transcriptase itself come from?

[@animatedbiologywitharpan]

You provide it in the reaction mixture, reverse transcrtase for this purpose is syntheticly made.

[@thisslightlysweetlife3402]

@@animatedbiologywitharpan In vivo I mean. Does it exist within our bodies or does the viral RNA code for it as well.

[@Apoorva4102]

@@thisslightlysweetlife3402 it's isolated from AMV virus :)

[@MolecularMedicine327]

Please refer the references i need to write my notes thanks in advance

[@animatedbiologywitharpan]

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[@user-sy5td7ft2u]

References please🙏

[@animatedbiologywitharpan]

www.thermofisher.com/in/en/home/life-science/pcr/reverse-transcription.html

[@Jesusandbible]

if you claim to be knowledgeable ...... is this a real quotation or not?
“Quantitative PCR is an oxymoron.” Kary Mullis, inventor of PCR.
As it it is then almost all the stats we have about Covid are bunk.