AlanFung:LabNotes/Methylome/2012-1-17: Difference between revisions
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==End repair & A-tailing== | ==End repair & A-tailing== | ||
===End Repair=== | |||
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| align="center" style="background:#f0f0f0;"|'''Content''' | | align="center" style="background:#f0f0f0;"|'''Content''' |
Revision as of 18:08, 20 January 2012
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
End Repair
Content | Volume |
Fragmented DNA | 50 |
End Reapir buffer | 10 |
End Repair Enzyme Mix | 5 |
ddH2O | 35 |
Volume | 100 |
- incubate at 20C for 30mins
- Purify DNA with Qiaquick column