Changing PCR recipe halfway - (Jun/03/2015 )

Hi all!

I have been using this PCR recipe because the company that makes the reverse transcriptase that I use recommends using 2uL of the cDNA in a 50uL reaction:

- 10uL 5x PCR mix

- 2.5uL 10uM forward primer + 2.5uL 10uM reverse primer

- 2uL cDNA

- 33uL ddH2O

But I feel like I'm running through the PCR mix awfully fast. So another recipe that I adapted the above one from is this 20uL recipe:

- 4uL 5x PCR mix

- 1uL 10uM forward primer + 1uL 10uM reverse primer

- 0.5uL cDNA

- 13.5uL ddH2O

If I switch from the 50uL recipe to 20uL recipe (and keep everything else the same), will my PCR results change?

Thank you!

Yes, it should do the trick.

Of course you can never predict things 100% its possible you have to adjust some stuff, but often it works.

There is only one way to find out: try it!