Noi/NOTES/2011-8-14
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BSPP DMR330k capture of brain sample from Dr. Ophoff's lab (UCLA)
- August 1, 2011, received 96 samples from Dr. Ophoff's lab
- Tina Wang confirmed the concentration of each sample was about 50ng/ul
- The concentrations were determined again by Nanadrop, and it turned out that the concentrations were vere varied. The volume of the gDNA needed to be adjust when perform bisulfite conversion
- Note: The ratio of 260/230nm of most of the samples were very low, so I assumed higj salt contamination.
Nanodrop measurement
Well ID | ng/ul | Well ID | ng/ul | Well ID | ng/ul | Well ID | ng/ul | Well ID | ng/ul | Well ID | ng/ul | Well ID | ng/ul | Well ID | ng/ul | Well ID | ng/ul | Well ID | ng/ul | Well ID | ng/ul | Well ID | ng/ul |
A1 | 127.53 | A2 | 101.77 | A3 | 104.16 | A4 | 89.42 | A5 | 108.53 | A6 | 98.50 | A7 | 71.86 | A8 | 59.88 | A9 | 76.42 | A10 | 59.93 | A11 | 152.47 | A12 | 74.20 |
B1 | 214.10 | B2 | 109.05 | B3 | 128.33 | B4 | 80.22 | B5 | 108.85 | B6 | 74.95 | B7 | 39.82 | B8 | 85.67 | B9 | 228.10 | B10 | 72.25 | B11 | 90.51 | B12 | 92.20 |
C1 | 98.39 | C2 | 106.42 | C3 | 90.52 | C4 | 114.26 | C5 | 100.28 | C6 | 91.90 | C7 | 79.72 | C8 | 96.65 | C9 | 119.19 | C10 | 69.85 | C11 | 106.38 | C12 | 46.85 |
D1 | 102.29 | D2 | 94.38 | D3 | 75.38 | D4 | 119.02 | D5 | 109.51 | D6 | 63.31 | D7 | 100.77 | D8 | 128.37 | D9 | 269.45 | D10 | 129.33 | D11 | 88.70 | D12 | 68.39 |
E1 | 90.70 | E2 | 84.67 | E3 | 139.81 | E4 | 90.16 | E5 | 98.92 | E6 | 76.35 | E7 | 197.06 | E8 | 89.01 | E9 | 49.91 | E10 | 62.05 | E11 | 78.12 | E12 | 83.77 |
F1 | 55.57 | F2 | 108.48 | F3 | 67.75 | F4 | 117.73 | F5 | 114.30 | F6 | 77.34 | F7 | 60.37 | F8 | 95.33 | F9 | 86.00 | F10 | 139.09 | F11 | 61.46 | F12 | 53.64 |
G1 | 111.01 | G2 | 62.01 | G3 | 98.83 | G4 | 133.36 | G5 | 166.66 | G6 | 100.09 | G7 | 53.66 | G8 | 84.04 | G9 | 593.63 | G10 | 189.64 | G11 | 111.77 | G12 | 106.40 |
H1 | 108.95 | H2 | 57.54 | H3 | 83.68 | H4 | 90.89 | H5 | 100.31 | H6 | 76.76 | H7 | 55.88 | H8 | 144.05 | H9 | 51.54 | H10 | 70.48 | H11 | 125.43 | H12 | 74.57 |
Bisulfite conversion
- Used EZ-96 DNA Methylation-Gold™ Kit (deep well format) Cat#D5008
Sample ID | ng/ul | Volume for 1200ng | Sample ID | H2O | Total amount (ng) | CT conversion reagent (ul) | Total volume (ul) |
A1 | 127.53 | 9.41 | A1 | 10.59 | 1200.00 | 130.00 | 150.00 |
B1 | 214.10 | 5.60 | B1 | 14.40 | 1200.00 | 130.00 | 150.00 |
C1 | 98.39 | 12.20 | C1 | 7.80 | 1200.00 | 130.00 | 150.00 |
D1 | 102.29 | 11.73 | D1 | 8.27 | 1200.00 | 130.00 | 150.00 |
E1 | 90.70 | 13.23 | E1 | 6.77 | 1200.00 | 130.00 | 150.00 |
F1* | 55.57 | 20.00 | F1* | 0.00 | 1111.40 | 130.00 | 150.00 |
G1 | 111.01 | 10.81 | G1 | 9.19 | 1200.00 | 130.00 | 150.00 |
H1 | 108.95 | 11.01 | H1 | 8.99 | 1200.00 | 130.00 | 150.00 |
A2 | 101.77 | 11.79 | A2 | 8.21 | 1200.00 | 130.00 | 150.00 |
B2 | 109.05 | 11.00 | B2 | 9.00 | 1200.00 | 130.00 | 150.00 |
C2 | 106.42 | 11.28 | C2 | 8.72 | 1200.00 | 130.00 | 150.00 |
D2 | 94.38 | 12.71 | D2 | 7.29 | 1200.00 | 130.00 | 150.00 |
E2 | 84.67 | 14.17 | E2 | 5.83 | 1200.00 | 130.00 | 150.00 |
F2 | 108.48 | 11.06 | F2 | 8.94 | 1200.00 | 130.00 | 150.00 |
G2 | 62.01 | 19.35 | G2 | 0.65 | 1200.00 | 130.00 | 150.00 |
H2* | 57.54 | 20.00 | H2* | 0.00 | 1150.80 | 130.00 | 150.00 |
A3 | 104.16 | 11.52 | A3 | 8.48 | 1200.00 | 130.00 | 150.00 |
B3 | 128.33 | 9.35 | B3 | 10.65 | 1200.00 | 130.00 | 150.00 |
C3 | 90.52 | 13.26 | C3 | 6.74 | 1200.00 | 130.00 | 150.00 |
D3 | 75.38 | 15.92 | D3 | 4.08 | 1200.00 | 130.00 | 150.00 |
E3 | 139.81 | 8.58 | E3 | 11.42 | 1200.00 | 130.00 | 150.00 |
F3 | 67.75 | 17.71 | F3 | 2.29 | 1200.00 | 130.00 | 150.00 |
G3 | 98.83 | 12.14 | G3 | 7.86 | 1200.00 | 130.00 | 150.00 |
H3 | 83.68 | 14.34 | H3 | 5.66 | 1200.00 | 130.00 | 150.00 |
A4 | 89.42 | 13.42 | A4 | 6.58 | 1200.00 | 130.00 | 150.00 |
B4 | 80.22 | 14.96 | B4 | 5.04 | 1200.00 | 130.00 | 150.00 |
C4 | 114.26 | 10.50 | C4 | 9.50 | 1200.00 | 130.00 | 150.00 |
D4 | 119.02 | 10.08 | D4 | 9.92 | 1200.00 | 130.00 | 150.00 |
E4 | 90.16 | 13.31 | E4 | 6.69 | 1200.00 | 130.00 | 150.00 |
F4 | 117.73 | 10.19 | F4 | 9.81 | 1200.00 | 130.00 | 150.00 |
G4 | 133.36 | 9.00 | G4 | 11.00 | 1200.00 | 130.00 | 150.00 |
H4 | 90.89 | 13.20 | H4 | 6.80 | 1200.00 | 130.00 | 150.00 |
A5 | 108.53 | 11.06 | A5 | 8.94 | 1200.00 | 130.00 | 150.00 |
B5 | 108.85 | 11.02 | B5 | 8.98 | 1200.00 | 130.00 | 150.00 |
C5 | 100.28 | 11.97 | C5 | 8.03 | 1200.00 | 130.00 | 150.00 |
D5 | 109.51 | 10.96 | D5 | 9.04 | 1200.00 | 130.00 | 150.00 |
E5 | 98.92 | 12.13 | E5 | 7.87 | 1200.00 | 130.00 | 150.00 |
F5 | 114.30 | 10.50 | F5 | 9.50 | 1200.00 | 130.00 | 150.00 |
G5 | 166.66 | 7.20 | G5 | 12.80 | 1200.00 | 130.00 | 150.00 |
H5 | 100.31 | 11.96 | H5 | 8.04 | 1200.00 | 130.00 | 150.00 |
A6 | 98.50 | 12.18 | A6 | 7.82 | 1200.00 | 130.00 | 150.00 |
B6 | 74.95 | 16.01 | B6 | 3.99 | 1200.00 | 130.00 | 150.00 |
C6 | 91.90 | 13.06 | C6 | 6.94 | 1200.00 | 130.00 | 150.00 |
D6 | 63.31 | 18.95 | D6 | 1.05 | 1200.00 | 130.00 | 150.00 |
E6 | 76.35 | 15.72 | E6 | 4.28 | 1200.00 | 130.00 | 150.00 |
F6 | 77.34 | 15.52 | F6 | 4.48 | 1200.00 | 130.00 | 150.00 |
G6 | 100.09 | 11.99 | G6 | 8.01 | 1200.00 | 130.00 | 150.00 |
H6 | 76.76 | 15.63 | H6 | 4.37 | 1200.00 | 130.00 | 150.00 |
A7 | 71.86 | 16.70 | A7 | 3.30 | 1200.00 | 130.00 | 150.00 |
B7** | 39.82 | 20.00 | B7** | 0.00 | 796.40 | 130.00 | 150.00 |
C7 | 79.72 | 15.05 | C7 | 4.95 | 1200.00 | 130.00 | 150.00 |
D7 | 100.77 | 11.91 | D7 | 8.09 | 1200.00 | 130.00 | 150.00 |
E7 | 197.06 | 6.09 | E7 | 13.91 | 1200.00 | 130.00 | 150.00 |
F7 | 60.37 | 19.88 | F7 | 0.12 | 1200.00 | 130.00 | 150.00 |
G7* | 53.66 | 20.00 | G7* | 0.00 | 1073.20 | 130.00 | 150.00 |
H8* | 55.88 | 20.00 | H8* | 0.00 | 1117.60 | 130.00 | 150.00 |
A8* | 59.88 | 20.00 | A8* | 0.00 | 1197.60 | 130.00 | 150.00 |
B8 | 85.67 | 14.01 | B8 | 5.99 | 1200.00 | 130.00 | 150.00 |
C8 | 96.65 | 12.42 | C8 | 7.58 | 1200.00 | 130.00 | 150.00 |
D8 | 128.37 | 9.35 | D8 | 10.65 | 1200.00 | 130.00 | 150.00 |
E8 | 89.01 | 13.48 | E8 | 6.52 | 1200.00 | 130.00 | 150.00 |
F8 | 95.33 | 12.59 | F8 | 7.41 | 1200.00 | 130.00 | 150.00 |
G8 | 84.04 | 14.28 | G8 | 5.72 | 1200.00 | 130.00 | 150.00 |
H8 | 144.05 | 8.33 | H8 | 11.67 | 1200.00 | 130.00 | 150.00 |
A9 | 76.42 | 15.70 | A9 | 4.30 | 1200.00 | 130.00 | 150.00 |
B9 | 228.10 | 5.26 | B9 | 14.74 | 1200.00 | 130.00 | 150.00 |
C9 | 119.19 | 10.07 | C9 | 9.93 | 1200.00 | 130.00 | 150.00 |
D9 | 269.45 | 4.45 | D9 | 15.55 | 1200.00 | 130.00 | 150.00 |
E9* | 49.91 | 20.00 | E9* | 0.00 | 998.20 | 130.00 | 150.00 |
F9 | 86.00 | 13.95 | F9 | 6.05 | 1200.00 | 130.00 | 150.00 |
G9*** | 593.63 | 2.02 | G9*** | 17.98 | 1200.00 | 130.00 | 150.00 |
H9* | 51.54 | 20.00 | H9* | 0.00 | 1030.80 | 130.00 | 150.00 |
A10 | 59.93 | 20.00 | A10 | 0.00 | 1198.60 | 130.00 | 150.00 |
B10 | 72.25 | 16.61 | B10 | 3.39 | 1200.00 | 130.00 | 150.00 |
C10 | 69.85 | 17.18 | C10 | 2.82 | 1200.00 | 130.00 | 150.00 |
D10 | 129.33 | 9.28 | D10 | 10.72 | 1200.00 | 130.00 | 150.00 |
E10 | 62.05 | 19.34 | E10 | 0.66 | 1200.00 | 130.00 | 150.00 |
F10 | 139.09 | 8.63 | F10 | 11.37 | 1200.00 | 130.00 | 150.00 |
G10 | 189.64 | 6.33 | G10 | 13.67 | 1200.00 | 130.00 | 150.00 |
H10 | 70.48 | 17.03 | H10 | 2.97 | 1200.00 | 130.00 | 150.00 |
A11 | 152.47 | 7.87 | A11 | 12.13 | 1200.00 | 130.00 | 150.00 |
B11 | 90.51 | 13.26 | B11 | 6.74 | 1200.00 | 130.00 | 150.00 |
C11 | 106.38 | 11.28 | C11 | 8.72 | 1200.00 | 130.00 | 150.00 |
D11 | 88.70 | 13.53 | D11 | 6.47 | 1200.00 | 130.00 | 150.00 |
E11 | 78.12 | 15.36 | E11 | 4.64 | 1200.00 | 130.00 | 150.00 |
F11 | 61.46 | 19.52 | F11 | 0.48 | 1200.00 | 130.00 | 150.00 |
G11 | 111.77 | 10.74 | G11 | 9.26 | 1200.00 | 130.00 | 150.00 |
H11 | 125.43 | 9.57 | H11 | 10.43 | 1200.00 | 130.00 | 150.00 |
A12 | 74.20 | 16.17 | A12 | 3.83 | 1200.00 | 130.00 | 150.00 |
B12 | 92.20 | 13.02 | B12 | 6.98 | 1200.00 | 130.00 | 150.00 |
C12** | 46.85 | 20.00 | C12* | 0.00 | 937.00 | 130.00 | 150.00 |
D12 | 68.39 | 17.55 | D12 | 2.45 | 1200.00 | 130.00 | 150.00 |
E12 | 83.77 | 14.32 | E12 | 5.68 | 1200.00 | 130.00 | 150.00 |
F12* | 53.64 | 20.00 | F12* | 0.00 | 1072.80 | 130.00 | 150.00 |
G12 | 106.40 | 11.28 | G12 | 8.72 | 1200.00 | 130.00 | 150.00 |
H12 | 74.57 | 16.09 | H12 | 3.91 | 1200.00 | 130.00 | 150.00 |
- Eluted volume 20ul with Elution buffer. After Qubit ss-DNA assay the total yields will be calculated based on ~16ul left in the well.
- Note:
- *= Total amount in bis-cvt lower than 1200ng, but still above 1000ng
- ** or ***= Performed two bis-cvt (one in the plate and one in the column (B7 (conc, too low, G9 (conc. too high)
- ***= conc. too high