Noi/NOTES/2012-5-25: Difference between revisions

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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

== 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.