Noi/NOTES/2012-5-25: Difference between revisions
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>Noi (Created page with "* Continued from:") |
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* Continued from: | * [[http://genome-tech.ucsd.edu/LabNotes/index.php/noi:DMR220k_LabNotes '''Link to calendar''']] | ||
* Continued from: http://genome-tech.ucsd.edu/LabNotes/index.php/Noi/NOTES/2012-5-21#SLN_mix_solution | |||
== PCR Amplification with barcoded primers (AmpF6.4Sol and AmpR6.3 IndX No.1-) | |||
* I tested if the capture works well by performing qPCR of few samples in small volume 25ul, sample E1, F1, G1 and NTC | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Captured template''' | |||
| align="center" style="background:#f0f0f0;"|'''2.5''' | |||
| align="center" style="background:#f0f0f0;"|'''0''' | |||
|- | |||
| 10uM AmpF6.4Sol||0.5||2.1 | |||
|- | |||
| 10uM AmpR6.3Sol||0.5||2.1 | |||
|- | |||
| 50x SYBG||0.2||0.84 | |||
|- | |||
| 2x Phusion MM||12.5||52.5 | |||
|- | |||
| H2O||8.8||36.96 | |||
|} | |||
* Aliquot 22.5ul to each tube and add 2.5ul of circularized DNA templates | |||
'''Program'''<br> | |||
98C 30s -> (98C 10s -> 58C 20s -> 72C 20s)x8 -> (98C 10s -> 72C 20s)x15 -> 72C 3min | |||
* Form the qPCR of tested samples, I could observe amplification in every samples except NTC, so I can continue amplify all samples and barcode with index primers. |
Revision as of 22:53, 25 May 2012
- [Link to calendar]
- Continued from: http://genome-tech.ucsd.edu/LabNotes/index.php/Noi/NOTES/2012-5-21#SLN_mix_solution
== PCR Amplification with barcoded primers (AmpF6.4Sol and AmpR6.3 IndX No.1-)
- I tested if the capture works well by performing qPCR of few samples in small volume 25ul, sample E1, F1, G1 and NTC
Captured template | 2.5 | 0 |
10uM AmpF6.4Sol | 0.5 | 2.1 |
10uM AmpR6.3Sol | 0.5 | 2.1 |
50x SYBG | 0.2 | 0.84 |
2x Phusion MM | 12.5 | 52.5 |
H2O | 8.8 | 36.96 |
- Aliquot 22.5ul to each tube and add 2.5ul of circularized DNA templates
Program
98C 30s -> (98C 10s -> 58C 20s -> 72C 20s)x8 -> (98C 10s -> 72C 20s)x15 -> 72C 3min
- Form the qPCR of tested samples, I could observe amplification in every samples except NTC, so I can continue amplify all samples and barcode with index primers.