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|| | |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|| | |09_2||NA21781*1 102307 ||240ng/ul x 1 tubes||8ul||12ul||130ul||65.6ng/ul x 20ul||2.19/2.91 | ||
|- | |- | ||
|09_3|| | |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|| | |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|| | |09_5||NA21731*1 101807 ||217.4ng/ul x 1 tube||10ul||10ul||130ul||8ng/ul x 10ul||1.37/1.01 | ||
|- | |- | ||
|09_6|| | |09_6||NA21846*1 100407 ||186.5ng/ul x 1 tube||11ul||9ul||130ul||14ng/ul x 20ul|| | ||
|- | |- | ||
|09_7|| | |09_7||NA21070*1 100406 ||329.7ng/ul x 1 tube||6ul||14ul||130ul||8ng/ul x 10ul||1.37/1.01 | ||
|- | |- | ||
|} | |} |
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