Noi/NOTES/2011-3-10
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[[1]]
DMR220k capture (Probe set S1-S3 plus Exp1-Exp3) and library construction of cell samples from Belmonte's lab[edit]
- Received 5 cell pellet samples from Belmonte's lab (1x10E6 cell/sample)
- Extracted gDNA using DNeasy Blood and Tissue kit (Qiagen, Cat# 69504)
- Elute with AE buffer (elute1: 200ul, elute2: 100ul)
Samples | No. of cell |
1. HGPS | 1.0E6 |
2. cHGPS1 | 1.0E6 |
3. cHGPS40 | 1.0E6 |
4. AWS | 1.0E6 |
5. cAWS1 | 1.0E6 |
- Note: 10ug of gDNA was aliquoted for Alice for exome sequencing (except cHGPS1 and AWS, 7ug each)
Bisulfite conversion[edit]
Samples | gDNA conc (ng/ul) | Total volume (ul) | Yields (ug) | Volume for bis-cvt | H2O | Total amount for bis-cvt (ng) | CT conversion reagent (ul) | Total volume (ul) |
1. HGPS | 75.70 | 300.00 | 22.71 | 16.00 | 4.00 | 1211.20 | 130.00 | 150.00 |
2. cHGPS1 | 57.10 | 300.00 | 17.13 | 20.00 | 0.00 | 1142.00 | 130.00 | 150.00 |
3. cHGPS40 | 100.20 | 300.00 | 30.06 | 12.00 | 8.00 | 1202.40 | 130.00 | 150.00 |
4. AWS | 42.20 | 300.00 | 12.66 | 20.00 | 0.00 | 844.00 | 130.00 | 150.00 |
5. cAWS1 | 130.10 | 300.00 | 39.03 | 9.22 | 10.78 | 1200.00 | 130.00 | 150.00 |
- follow standard protocol for bisulfite conversion of ZymoResearch
- elute with 17ul Elution buffer
- Keep in -20C, waiting for Qubit ssDNA assay kit
- Continued on 2011_03_18: [[2]]