View Single Post
Old 08-17-2017, 08:10 PM   #8
Jafar Jabbari
Location: Melbourne

Join Date: Jan 2013
Posts: 1,196

It seems OK. Final DNA recovery from Pippin depends on the quantity of fragments in the selected size range which will vary based on input, genome composition and restriction enzymes used among other factors.

I think you can safely set up four 20 ul reactions using 5 ul of size selected tags per reaction and do 12 cycles. After 12 cycle you can check the PCR yield and size by TapeStation if it is available or at least check concentration with Qubit or PicoGreen. If PCR yield is around 1 ug/ul then no extra cycles would be needed. If it is lower then do 1 or 2 more cycles considering that each cycle yield will double.
nucacidhunter is offline   Reply With Quote