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]]