Jie:LabNotes/CpgSeq/2009-5-20: Difference between revisions

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| align="center" style="background:#f0f0f0;"|'''conversion reagents'''
| align="center" style="background:#f0f0f0;"|'''conversion reagents'''
| align="center" style="background:#f0f0f0;"|'''conversed DNA concentration and volumn'''
| align="center" style="background:#f0f0f0;"|'''conversed DNA concentration and volumn'''
| align="center" style="background:#f0f0f0;"|'''260:280/260:230'''
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|09_1||NA21687*1 062507 ||241.1ng/ul x 1 tubes||8ul||12ul||130ul||43.3ng/ul x 20ul||2.18/2.97
|09_1||NA21687*1 062507 ||241.1ng/ul x 1 tubes||8ul||12ul||130ul||95.4ng/ul x 10ul
|-
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|09_2||NA21781*1 102307 ||240ng/ul x 1 tubes||8ul||12ul||130ul||65.6ng/ul x 20ul||2.19/2.91
|09_2||NA21781*1 102307 ||240ng/ul x 1 tubes||8ul||12ul||130ul||120.2ng/ul x 10ul
|-
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|09_3||NA21833*1 101807 ||253.9ng/ul x 1 tube||8ul||12ul||130ul||22.7ng/ul x 10ul||2.05/2.78
|09_3||NA21833*1 101807 ||253.9ng/ul x 1 tube||8ul||12ul||130ul||121.5ng/ul x 10ul
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|09_4||NA21660*1 053007 ||243.2ng/ul x 1 tube||8ul||12ul||130ul||174.5ng/ul x 10ul||2.3/3.0
|09_4||NA21660*1 053007 ||243.2ng/ul x 1 tube||8ul||12ul||130ul||120.3ng/ul x 10ul
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|09_5||NA21731*1 101807 ||217.4ng/ul x 1 tube||10ul||10ul||130ul||8ng/ul x 10ul||1.37/1.01
|09_5||NA21731*1 101807 ||217.4ng/ul x 1 tube||10ul||10ul||130ul||127.3ng/ul x 10ul
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|09_6||NA21846*1 100407 ||186.5ng/ul x 1 tube||11ul||9ul||130ul||14ng/ul x 20ul||
|09_6||NA21846*1 100407 ||186.5ng/ul x 1 tube||11ul||9ul||130ul||151.3ng/ul x 10ul
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|09_7||NA21070*1 100406 ||329.7ng/ul x 1 tube||6ul||14ul||130ul||8ng/ul x 10ul||1.37/1.01
|09_7||NA21070*1 100406 ||329.7ng/ul x 1 tube||6ul||14ul||130ul||311.2ng/ul x 10ul
|-
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|||Jurkat gDNA ||140ng/ul x 3 tube||14ul||6ul||130ul||8ng/ul x 10ul||1.37/1.01
|||Jurkat gDNA ||140ng/ul x 3 tube||14ul||6ul||130ul||255.6ng/ul x 20ul
|-
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|}
|}

Revision as of 01:41, 22 May 2009

bisulfite conversion of PGP samples

sample sample concentration sample volumn ddH2O conversion reagents conversed DNA concentration and volumn
09_1 NA21687*1 062507 241.1ng/ul x 1 tubes 8ul 12ul 130ul 95.4ng/ul x 10ul
09_2 NA21781*1 102307 240ng/ul x 1 tubes 8ul 12ul 130ul 120.2ng/ul x 10ul
09_3 NA21833*1 101807 253.9ng/ul x 1 tube 8ul 12ul 130ul 121.5ng/ul x 10ul
09_4 NA21660*1 053007 243.2ng/ul x 1 tube 8ul 12ul 130ul 120.3ng/ul x 10ul
09_5 NA21731*1 101807 217.4ng/ul x 1 tube 10ul 10ul 130ul 127.3ng/ul x 10ul
09_6 NA21846*1 100407 186.5ng/ul x 1 tube 11ul 9ul 130ul 151.3ng/ul x 10ul
09_7 NA21070*1 100406 329.7ng/ul x 1 tube 6ul 14ul 130ul 311.2ng/ul x 10ul
Jurkat gDNA 140ng/ul x 3 tube 14ul 6ul 130ul 255.6ng/ul x 20ul


  • Add 900ul ddH2O, 300ul M-Dilution Buffer and 50ul M-Dissolving Buffer to a tube of CT Conversion Reagent.
  • Add 130ul of CT conversion reagent to samples.
  • 98C 10min -> 64C 150min -> 4c overnight.
  • Add 600ul M-Binding buffer to spin columns.
  • Add the converted samples to the columns, close the gap and mix by inverting several times.
  • Spin at 15,000rpm for 30sec.
  • Add 100ul M-Wash Buffer, spin for 30 sec, discard flow-through.
  • Add 200ul M-Desulphonation Buffer, wait for 20min, spin at 15,000 rpm for 30sec.
  • Add 200ul M-Wash Buffer, spin for 30 sec.
  • Place the columns into 1.5ml tubes, add 10ul M-Elution Buffer. Wait for 1min, spin at 15,000rpm for 30sec.
  • Measure the DNA with Nanodrop:

RNA