Daniel:Notebook/HiResChrPaint/2014-8-18: Difference between revisions
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>Djacobse (→qPCR) |
>Djacobse |
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##72C 2 min | ##72C 2 min | ||
##Hold at 16C forever | ##Hold at 16C forever | ||
#Ethanol Precipitation | #[[Daniel:Protocols/EtOHPrecip|Ethanol Precipitation]] | ||
##Continued [[Daniel:Notebook/HiResChrPaint/2014-8-19|tomorrow]] | |||
===qPCR Results=== | ===qPCR Results=== | ||
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[[Image:2014-08-18_PPRL.png|Successful run on all lanes|500px]] | [[Image:2014-08-18_PPRL.png|Successful run on all lanes|500px]] | ||
[[Category:HRCP]] [[Category:qPCR]] [[Category:20140813]] | |||
Latest revision as of 22:38, 9 September 2014
Padlock Probes (Started 8/13/2014)[edit]
qPCR[edit]
I need more padlock probes, so I'm going to repeat qPCR with 24x samples.
Reaction Table
Component | 1X Reaction | 12.2X Scaled |
Padlock Probes | 0.2 | 2.44 |
100 uM pAP1V41U | 0.4 | 4.88 |
100 uM pAP2V4 | 0.4 | 4.88 |
KAPA SYBR Fast Master Mix (2X) | 50 | 610 |
H2O | 49 | 597.8 |
Total | 100 | 1220 |
- Prepare master mix according to table above
- Add 100 uL reaction to PCR tubes (5 total)
- qPCR Protocol
- 95C 30 sec
- 95C 10 sec
- 55C 20 sec
- 72C 30 sec
- goto (3.2) 17 times
- 72C 2 min
- Hold at 16C forever
- Ethanol Precipitation
- Continued tomorrow