Hi all,
I am trying to prepare RRBS-seq library, but have some questions regarding the MspI digestion. I find my results a bit odd, since not much is digested and I have not been able to find a representative digestion of gDNA with MspI.
I did a time course (1hr, 2hrs, 6hrs and ON) of 1ug gDNA digested with 20U MspI from NEB, but the shift is not great.
Is the enzyme not working or is this to be expected, since RRBS is only taking 5-10% of the genome into consideration. How come that I also already see a bit of degradation in the lane with undigested gDNA?
Thanks a lot. Appreciate your input.
// Kevin
I am trying to prepare RRBS-seq library, but have some questions regarding the MspI digestion. I find my results a bit odd, since not much is digested and I have not been able to find a representative digestion of gDNA with MspI.
I did a time course (1hr, 2hrs, 6hrs and ON) of 1ug gDNA digested with 20U MspI from NEB, but the shift is not great.
Is the enzyme not working or is this to be expected, since RRBS is only taking 5-10% of the genome into consideration. How come that I also already see a bit of degradation in the lane with undigested gDNA?
Thanks a lot. Appreciate your input.
// Kevin
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