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

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(New page: ==bisulfite conversion of PGP samples== {| border="1" cellpadding="5" cellspacing="0" align="center" |- | align="center" style="background:#f0f0f0;"|''' ''' | align="center" style="backgr...)
 
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| align="center" style="background:#f0f0f0;"|'''260:280/260:230'''
| align="center" style="background:#f0f0f0;"|'''260:280/260:230'''
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|09_1||BJ||178ng/ul x 2 tubes||11.2ul||8.8ul||130ul||43.3ng/ul x 20ul||2.18/2.97
|09_1||NA21687*1 062507 ||241.1ng/ul x 1 tubes||8ul||12ul||130ul||43.3ng/ul x 20ul||2.18/2.97
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|09_2||BJ_iPS_11||140ng/ul x 2 tubes||14.3ul||5.7ul||130ul||65.6ng/ul x 20ul||2.19/2.91
|09_2||NA21781*1 102307 ||240ng/ul x 1 tubes||8ul||12ul||130ul||65.6ng/ul x 20ul||2.19/2.91
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|09_3||IMB90||50ng/ul||20ul||0ul||130ul||22.7ng/ul x 10ul||2.05/2.78
|09_3||NA21833*1 101807 ||253.9ng/ul x 1 tube||8ul||12ul||130ul||22.7ng/ul x 10ul||2.05/2.78
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|09_4||Hybrid2||200ng/ul||10ul||10ul||130ul||174.5ng/ul x 10ul||2.3/3.0
|09_4||NA21660*1 053007 ||243.2ng/ul x 1 tube||8ul||12ul||130ul||174.5ng/ul x 10ul||2.3/3.0
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|09_5||Hues8||178.5ng/ul||13ul||7ul||130ul||8ng/ul x 10ul||1.37/1.01
|09_5||NA21731*1 101807 ||217.4ng/ul x 1 tube||10ul||10ul||130ul||8ng/ul x 10ul||1.37/1.01
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|09_6||Hues42||20ng/ul x 2 tube||20ul||0ul||130ul||14ng/ul x 20ul||
|09_6||NA21846*1 100407 ||186.5ng/ul x 1 tube||11ul||9ul||130ul||14ng/ul x 20ul||
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|09_7||Hues8||178.5ng/ul||13ul||7ul||130ul||8ng/ul x 10ul||1.37/1.01
|09_7||NA21070*1 100406 ||329.7ng/ul x 1 tube||6ul||14ul||130ul||8ng/ul x 10ul||1.37/1.01
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Revision as of 22:57, 20 May 2009

bisulfite conversion of PGP samples

sample sample concentration sample volumn ddH2O conversion reagents conversed DNA concentration and volumn 260:280/260:230
09_1 NA21687*1 062507 241.1ng/ul x 1 tubes 8ul 12ul 130ul 43.3ng/ul x 20ul 2.18/2.97
09_2 NA21781*1 102307 240ng/ul x 1 tubes 8ul 12ul 130ul 65.6ng/ul x 20ul 2.19/2.91
09_3 NA21833*1 101807 253.9ng/ul x 1 tube 8ul 12ul 130ul 22.7ng/ul x 10ul 2.05/2.78
09_4 NA21660*1 053007 243.2ng/ul x 1 tube 8ul 12ul 130ul 174.5ng/ul x 10ul 2.3/3.0
09_5 NA21731*1 101807 217.4ng/ul x 1 tube 10ul 10ul 130ul 8ng/ul x 10ul 1.37/1.01
09_6 NA21846*1 100407 186.5ng/ul x 1 tube 11ul 9ul 130ul 14ng/ul x 20ul
09_7 NA21070*1 100406 329.7ng/ul x 1 tube 6ul 14ul 130ul 8ng/ul x 10ul 1.37/1.01


  • 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