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Posted
Real Time PCR or Reverse Transcription PCR?

Uh, i never thought that they are different things. :eek: I would test up or down regulation of gene expression. Which one should i use?

 

THanks. :)

Posted

Both Real time PCR and Reverse transcription pcr are capable of doing quantitative analysis of mRNA. Real time is alot more accurate and quicker once setup, but more technically tricky to set up. Beacon design n all.

  • 3 months later...
Posted

Although (unfortunately) some people use the term RT-PCR for real-time PCR, the most commonly accepted, old and widespread meaning for RT is reverse transcription (or reverse transcriptase).

 

If you are talking about gene expression, you need a reverse transcription step. This creates molecules of DNA (cDNA or complementary DNA) from mRNA molecules (messenger RNA). The cDNA is subsequently used in a PCR reaction (hence the name RT-PCR).

 

As regards to the other meaning (real-time PCR) is not necessary related to the study of gene expression. It's a technique that allows you to determine the amount of template DNA that is present at the beginning of the PCR. This template can be cDNA (after a reverse transcription step, to quantify expression levels), but can also be viral or bacterial DNA (to assess the progress of an infection), genomic DNA (to assess the number of transgenes in a transgenic plant), etc.

 

Hope this helps. :cool:

  • 1 month later...
Posted
Although (unfortunately) some people use the term RT-PCR for real-time PCR' date=' the most commonly accepted, old and widespread meaning for RT is reverse transcription (or reverse transcriptase).

 

If you are talking about gene expression, you need a reverse transcription step. This creates molecules of DNA (cDNA or complementary DNA) from mRNA molecules (messenger RNA). The cDNA is subsequently used in a PCR reaction (hence the name RT-PCR).

 

As regards to the other meaning (real-time PCR) is not necessary related to the study of gene expression. It's a technique that allows you to determine the amount of template DNA that is present at the beginning of the PCR. This template can be cDNA (after a reverse transcription step, to quantify expression levels), but can also be viral or bacterial DNA (to assess the progress of an infection), genomic DNA (to assess the number of transgenes in a transgenic plant), etc.

 

Hope this helps. :cool:[/quote']

 

And of course, you have to do an RT in order to do real-time...

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