PCR product size 1.2 kb optimum yield after 30 cycle 20 ul reaction
The simplest answer is to determine molecular weight of a 1.2 kb DNA and convert that to moles and weight. A 1.2 kb DNA will have a molecular weight of 1200 X 660 grams per mole (6.02 X 10^23 molecules). One DNA fragment will yield 2^30 copies under ideal (100% efficiency) PCR conditions. Factor in the amount of templates you start with for your PCR, make some assumptions on PCR efficiency and you can come up with an estimate. My calculations show that one 1.2 kb DNA fragment will yield about 1.41 ng material after 30 PCR cycles at 100% efficiency. My experience is that the yield for a 1 kb fragment after 35 cycles is in the 5-6 ug range with 25 ng starting template.
I use pcr for gene synthesis so I need good yield I ask whats avarage yield I could get after each reaction
The simplest answer is to determine molecular weight of a 1.2 kb DNA and convert that to moles and weight. A 1.2 kb DNA will have a molecular weight of 1200 X 660 grams per mole (6.02 X 10^23 molecules). One DNA fragment will yield 2^30 copies under ideal (100% efficiency) PCR conditions. Factor in the amount of templates you start with for your PCR, make some assumptions on PCR efficiency and you can come up with an estimate. My calculations show that one 1.2 kb DNA fragment will yield about 1.41 ng material after 30 PCR cycles at 100% efficiency. My experience is that the yield for a 1 kb fragment after 35 cycles is in the 5-6 ug range with 25 ng starting template.
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