Noi/NOTES/2012-5-24
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DMR 330k BSPP capture of samples from Harvard's group, ASM and ASCM in human cells project
- Will be using gDNA sent by Anwesha on 2012_05_24
Sample IDs | Conc. (ng.ul) by Nanodrop (from Harvard) |
Amount (ug) in 40ul |
GM gDNA 4-5-12 | 64.52 | 2.58 |
P10E3 gDNA 4-5-12 | 37.86 | 1.51 |
DF2 gDNA 4-5-12 | 70.00 | 2.80 |
H0 gDNA 4-5-12 | 84.52 | 3.38 |
H7 gDNA 4-5-12 | 184.33 | 7.37 |
H14 gDNA 4-5-12 | 167.18 | 6.69 |
H16 gDNA 4-5-12 | 120.60 | 4.82 |
- After confirming concentration by Nanodrop, the conc. I got pretty close to the conc. they labeled.
Bisulfite conversion
Reagent prep Add the following reagents to CT Conversion Reagent (1tube) - H2O 900ul - M-Dissolving Buffer 50ul - M-Dilution Buffer 300ul Mix at room temperature with frequent vortexing or shaking for 15 minutes
Sample IDs | Conc. (ng/ul) | Volume for 1200ng | LabID ID | H2O | Total amount (ng) | CT conversion reagent (ul) | Total volume (ul) |
GM gDNA 4-5-12 | 64.52 | 18.60 | GM | 1.40 | 1200.00 | 130.00 | 150.00 |
P10E3 gDNA 4-5-12 | 37.86 | 20.00 | P10E | 0.00 | 757.20 | 130.00 | 150.00 |
DF2 gDNA 4-5-12 | 70.00 | 17.14 | DF2 | 2.86 | 1200.00 | 130.00 | 150.00 |
H0 gDNA 4-5-12 | 84.52 | 14.20 | H0 | 5.80 | 1200.00 | 130.00 | 150.00 |
H7 gDNA 4-5-12 | 184.33 | 6.51 | H7 | 13.49 | 1200.00 | 130.00 | 150.00 |
H14 gDNA 4-5-12 | 167.18 | 7.18 | H14 | 12.82 | 1200.00 | 130.00 | 150.00 |
H16 gDNA 4-5-12 | 120.60 | 9.95 | H16 | 10.05 | 1200.00 | 130.00 | 150.00 |
GM+H0:: 1:1 | 9.3 (GM)+ 7.1 (H0) | GM+H0 | 3.60 | 1200.13 | 130.00 | 150.00 |
Program 98°C for 10 minutes 64°C for 2.5 hours 4°C storage up to 20 hours Measure conc. with ssDNA Qubit assay (used 1ul of bis-cvt gDNA)