silly control question
Posted 13 March 2010 - 02:45 AM
I am quite new to PCR but there is something I donít understand
Why is it that when I use a No RT ( a sample that hasnít gone through reverse transcription) as a control I see no amplification - I am aware this is a good thing. But basically why doesnít the mRNA act as a template and produce no product? after all I have primers and polymerase in there that would recognise it? Is it to do with the U bases preventing primer annealing? But this cannot just be the only thing preventing this amplification. Once the mRNA is turned to cDNA everything appears to be fine.
Posted 14 March 2010 - 01:35 AM
Posted 17 March 2010 - 03:55 AM
To be precise, it can only catalyse polymerisation of DNA with DNA as a template. It doesn't recognise RNA.
Your polymerase is a DNA polymerase, and can therefore only catalyse the polymerisation of DNA and not RNA.
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