Noi/NOTES/2012-7-27

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Revision as of 15:46, 27 July 2012 by >Noi (Created page with "http://genome-tech.ucsd.edu/LabNotes/index.php/noi:DMR220k_LabNotes '''Link to calendar'''<br> = WGBS sequencing library preparation using KAPA protocol (Eppendorf) = * No...")
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WGBS sequencing library preparation using KAPA protocol (Eppendorf)

  • Note that I used the same total volume of reaction similar to the protocol from Eppendorf. This might be slightly different from the original protocol from KAPA kit, but the total amount of enzyme and buffer are the same. I also adjust the volume of adapters in ligation step, but total volume and ratio of adapter to A-tailed DNA are the same.

Preparation of enzyme mix for End repair, A-tailing, Ligation, and Methylation adapters

  • First of all, need to check if I have enough reagents for 8 rxn
  • Since I perform library prep for exactly 8 rxn (1 column-> single pipetting), I will not prepare extra volume since I plane to set up machine to pipette all reaction mix aliquoted in the well (exactly each reaction need). This mean setting up larger volume of pipetting to make sure that all reaction mix is transfered to the DNA.