Noi/NOTES/2014-10-22
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Preparation of WGBS of cfDNA from plasma using KAPA Hyper Prep kit
- Rational: Will be added
Sample list
No. Sample IDs Conc. (ng/ul) #1 NC-2 0.325 #2 NC-3 0.538 #3 6P-2 1.10 #4 6P-3 1.57
- Note: I will reduce the reaction volume to be half of the instruction and reduce the reagent volume/amount proportionally.
- Actual reaction set up:
- - 50ul of fragmented DNA
- - 7ul of End repair & dA-tailing buffer
- - 3ul of End repair & dA-tailing enzyme mix
- - Total reaction: 60ul
End repair & dA-tailing
End repair & dA-tailing reaction mix
Components | Volume | half rxn | 5.2x rxn |
End repair/dA-tailing buffer | 7.00 | 3.50 | 18.20 |
End repair/dA-tailing enzyme mix | 3.00 | 1.50 | 7.80 |
Total | 10.00 | 5.00 |
Reaction set up
Sample IDs | Conc. (ng/ul) | Volume for 1ng (ul) | End repair/dA-tailing rxn mix | H2O (ul) | Total (ul) |
NC-2 | 0.33 | 3.08 | 5.00 | 21.92 | 30.00 |
NC-3 | 0.54 | 1.86 | 5.00 | 23.14 | 30.00 |
6P-2 | 1.10 | 0.91 | 5.00 | 24.09 | 30.00 |
6P-3 | 1.57 | 0.64 | 5.00 | 24.36 | 30.00 |
NTC | 0.00 | 0.00 | 5.00 | 25.00 | 30.00 |
- - Mix the reaction mix well
- - 20C, 30min
- - 65C, 30min,
- - Hold 4C
- - Proceed to Adaptor ligation immediately