We are doing Illumina libraries for Chip-seq with great success - however, we would want to know if anyone ever tried to reduce the PCR volume for the final amplification.
We usually have very low starting material and minimize the number of PCR cycles we do. We think that with 50 uL PCR reaction (KAPA Hifi) we might be wasting a lot of money as saturation is never reached due to low starting material and low number of cycles.
Anyone tried with 25uL or even 10uL? We are about to do a lot of libraries and it would be nice to know if this has ever been tried.
Thanks!
We usually have very low starting material and minimize the number of PCR cycles we do. We think that with 50 uL PCR reaction (KAPA Hifi) we might be wasting a lot of money as saturation is never reached due to low starting material and low number of cycles.
Anyone tried with 25uL or even 10uL? We are about to do a lot of libraries and it would be nice to know if this has ever been tried.
Thanks!