AlanFung:LabNotes/Methylome/2012-1-17

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Revision as of 23:53, 19 January 2012 by >Alan6017518 (→‎End repair & A-tailing)
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GM12878 Library

Sample Preparation

  • Get GM12878 samples from Noi
  • Measure concentration with qubit
  • Take 5ug of samples and prepare for shearing (Make total volume to 100ul with ddh2o)
  • Covaris shearing of 5ug gDNA to ~400bp
  • Contacted Yang at Kelly Frazer's lab to help me with it (so Yang says they don't have the coavris shearing machine anymore and they shear their sampels at Biogem now, emailed James and see if i can get it sheared over there)
  • James helped me out to shear the samples, he used the following settings

End repair & A-tailing

Content Volume
Fragmented DNA 50
End Reapir buffer 10
End Repair Enzyme Mix 5
ddH2O 35
Volume 100

ligation to methylated Y adaptor

Bisulfite Conversion

PCR

Size selection