need to know minimum amount of template DNA needed for pcr amplification
Posted 28 November 2010 - 06:42 PM
I am new to this forum ...I am currently using a Qiagen's hotstar Taq Mastermix for my PCR reaction. My question is what is the miminum amount of template DNA beyond which there will be Undetectable amplification product ?
Any feedback would be great !
Posted 01 December 2010 - 09:21 PM
-a new technique called digital PCR tries to utilize the point which the dilution of template is so low that some number of wells won't amplify. if you look up the literature on that technique i suspect you can calculate the theoretical point at which their is likely to be less than 1 copy your template in the reaction
-if you are trying to get by on really low amounts of template, you might try increasing your number of cycles, or even doing a nested PCR
-you could just test this empirically by creating a standard curve
Posted 02 December 2010 - 04:43 AM
One must presume that long and short arguments contribute to the same end. - Epicurus
...except casandra's that belong to the funniest, most interesting and imaginative (or over-imaginative?) ones, I suppose.