asymmetric PCR (View forum version)


Posted 11 November 2009 - 07:03 AM

Hi there!
I've heard that I can carry out an asymmetric PCR in order to have a ssDNA, is it right? And then, could anybody suggest a protocol to carry it out?



Posted 11 November 2009 - 02:14 PM

You simply need to add a single primer rather than two primers in your pcr reaction. You do need to be aware that the amplification will be linear rather than exponential in the number of cycles, so it will matter a great deal how much initial template you use. If you do 35 cycles, you will only get 35 times as much ssDNA as you had initial dsDNA template.