Jie:LabNotes/CpgSeq/2008-10-10
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Bisulfite conversion of DNA using Zymo EZ DNA Methylation-Gold kit
sample | sample concentration | sample volumn | ddH2O | conversion reagents | conversed DNA concentration and volumn | 260:280/260:230 | |||||||||||||||||||||||||||||||||||||
2 | BJ_iPS_11 | 140ng/ul x 2 tubes | 14.3ul | 5.7ul | 130ul | 65.6ng/ul x 20ul | 2.19/2.91 | ||||||||||||||||||||||||||||||||||||
16 | IMB90_2 | 50ng/ul | 20ul | 0ul | 130ul | 22.7ng/ul x 10ul | 2.05/2.78 | ||||||||||||||||||||||||||||||||||||
17 |
RNA set up the capture system
dNTP concentration 10mM(NEB) ddH2O Ligase Buffer Ligase Amplitaq dNTP 2U/ul AmpliTaq Stoffel fragment; 0.5U/ul AmpLigase; 0.2mM dNTP: 0.58ul 0.1ul 0.1ul 0.2ul (10mM)0.02ul x300 174ul 30ul 30ul 60ul 6ul 95c 10min -> 55C 16h ->add 1ul SLN mix(2U/ul AmpliTaq Stoffel fragment; 0.5U/ul AmpLigase; 0.2mM dNTP) -> 55C 4h -> |