AlanFung:LabNotes/2014/2014-9-8

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Revision as of 23:24, 9 September 2014 by >Alan6017518 (→‎qPCR Test to verifiy End repair, A-tailing & Adapter Ligation)
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qPCR Test to verifiy End repair, A-tailing & Adapter Ligation

Component 1 Library 19 Libraries + 3%
qPCR Master Mix:    
Adapter ligated DNA 1 19.6
10uM PCR_F 0.5 9.8
10uM PCR_R.Ind2 0.5 9.8
2X Kapa Fast 12.5 244.6
Water 10.5 205.5
Total master mix volume 25 489.3
  • 98C 2min --> (98C 15sec -> 64C 20sec -> 72C 30sec) x 12 cycles --> 72C 3min --> 15C Hold

File:09082014 Cycle12.JPG

PAGE Gel Verification

File:ZhangLab 2 2014-09-08 16hr 52min.jpg

  • Bands ~110bp is most likely primer dimers, it was not removed by the beads as 1.7X beads will only remove anything below 100bp
  • pick a few samples and Repeat reaction with 7,8,9 cycle
  • Run PAGE gel with negative control and