AlanFung:LabNotes/CTCF/2010-6-7: Difference between revisions
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>Alan6017518 |
>Alan6017518 |
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*Qiaquick Purification | *Qiaquick Purification | ||
*Nanodrop | *Nanodrop | ||
P7-6.4ng/ul | |||
P10-5.4ng/ul | |||
*Gel Quant | *Gel Quant | ||
[[File:ZhangLab_2 2010-06-07 14hr 34min.jpg]] | [[File:ZhangLab_2 2010-06-07 14hr 34min.jpg]] | ||
===Protocol=== | |||
Obtain 100ng Jurkat DNA | |||
*100ng/(6.4ng/ul)=15.63ul | |||
*100ng/(5.4ng/ul)=18.52ul | |||
{| {{table}} | |||
| ||P7||P10 | |||
|- | |||
| Jurkat DNA PCR Product||15.63||18.52 | |||
|- | |||
| ddH2O||69.37||66.48 | |||
|- | |||
| End Repair Reaction Buffer (10x)||10||10 | |||
|- | |||
| Repair Enzyme Mix||5||5 | |||
|- | |||
| | |||
|} | |||
*Incubate 30 mins at 20C | |||
Purification using qiaquick column | |||
*Elute with 50ul buffer | |||
*Nanodrop result - 6.5ng/ul*50ul |
Revision as of 21:55, 7 June 2010
Prepare PCR Product from Jurkat gDNA
ul | |
primer | 1 |
h2o | 7 |
template | 2 |
Econo 2x | 10 |
- Program: 94C 2min-> (94C 40sec-> 52C 40sec -> 72C 60 sec) x 2 -> 72C 10min -> 15C hold
- Qiaquick Purification
- Nanodrop
P7-6.4ng/ul P10-5.4ng/ul
- Gel Quant
File:ZhangLab 2 2010-06-07 14hr 34min.jpg
Protocol
Obtain 100ng Jurkat DNA
- 100ng/(6.4ng/ul)=15.63ul
- 100ng/(5.4ng/ul)=18.52ul
P7 | P10 | |
Jurkat DNA PCR Product | 15.63 | 18.52 |
ddH2O | 69.37 | 66.48 |
End Repair Reaction Buffer (10x) | 10 | 10 |
Repair Enzyme Mix | 5 | 5 |
- Incubate 30 mins at 20C
Purification using qiaquick column
- Elute with 50ul buffer
- Nanodrop result - 6.5ng/ul*50ul