[PCR] What if I do a PCR with 99 cycles...

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bbs_r
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[PCR] What if I do a PCR with 99 cycles...

Post by bbs_r » Wed Mar 29, 2006 2:09 am

I am just curious with the fact that the more the cycles I run in the PCR, the more the products I will get.

So, my question is:
If I told the PCR machine to run 99 cycles, does there any negative effects apart from time wasting?

Thank you very much!

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LilKim
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Post by LilKim » Wed Mar 29, 2006 3:07 am

PCR has exponential amplification.... however the curve "flattens" because primers and DNTP will limit any standard reaction. (they both become incorporated into the amplified products so.. you'll run-out eventually)

... I dunno if you can get around this if you added more?? but again, this would throw-off concentrations of things in the mix...

this is just my guess anyways...

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canalon
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Post by canalon » Wed Mar 29, 2006 4:09 am

Yep and excess nucleotides inhibits PCR from the start as I recently experimented while changing lab (and dNTPs mixes, while keeping old habits). So you'll get better results by simply preparing multiple tubes tahn by increasing the number of cycles.
Patrick

Science has proof without any certainty. Creationists have certainty without
any proof. (Ashley Montague)

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