Daniel:Notebook/HiResChrPaint/2014-1-8: Difference between revisions
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>Djacobse (Created page with "=Alu Control= Back to Calendar Following the results from December 18, it appears that my prim...") |
>Djacobse |
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1. Reaction table | 1. Reaction table | ||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | |||
|- style="font-size:12pt" align="center" | |||
|style="background-color:#FFFF00" align="center" width="65" height="45" | | |||
|style="background-color:#D9D9D9;font-weight:bold" width="65" | gDNA | |||
|style="background-color:#D9D9D9;font-weight:bold" width="65" | 2X Kapa SYBR qPCR MM | |||
|style="background-color:#D9D9D9;font-weight:bold" width="65" | 100 uM AP1V61U-Amino | |||
|style="background-color:#D9D9D9;font-weight:bold" width="65" | 100 uM AP2V6 | |||
|style="background-color:#D9D9D9;font-weight:bold" width="65" | H20 | |||
|style="background-color:#D9D9D9;font-weight:bold" width="65" | Total Volume | |||
|- style="background-color:#8DB4E2;font-size:12pt" align="center" | |||
| height="15" | per rxn | |||
| align="center" | 1 | |||
| align="center" | 50 | |||
| align="center" | 0.5 | |||
| align="center" | 0.5 | |||
| align="center" | 48 | |||
| align="center" | 100 | |||
|- style="font-size:12pt" align="center" | |||
|style="background-color:#D9D9D9;font-weight:bold" height="30" | PGP1F-A (12.1X) | |||
| align="center" | 12.1 | |||
| align="center" | 605 | |||
| align="center" | 6.05 | |||
| align="center" | 6.05 | |||
| align="center" | 580.8 | |||
| align="center" | 1210 | |||
|} | |||
2. Aliquot 100 uL into each of 3 qPCR tubes | 2. Aliquot 100 uL into each of 3 qPCR tubes |
Revision as of 23:48, 7 January 2014
Alu Control
Following the results from December 18, it appears that my primers work. However, I used up all the sample from the previous primary amplification, so I'll have to re-amplify from genomic DNA (see Genomic DNA Extraction Results).
Expansion qPCR
1. Reaction table
gDNA | 2X Kapa SYBR qPCR MM | 100 uM AP1V61U-Amino | 100 uM AP2V6 | H20 | Total Volume | |
per rxn | 1 | 50 | 0.5 | 0.5 | 48 | 100 |
PGP1F-A (12.1X) | 12.1 | 605 | 6.05 | 6.05 | 580.8 | 1210 |
2. Aliquot 100 uL into each of 3 qPCR tubes 3. PCR protocol: i. PCR 3 min 95C ii. 95C 10 sec iii. 55C 30 sec iv. 72C 20 sec v. plate read vi. goto ii x17 vii. 72C 2 min viii. 16C hold 4. Pool 3 samples together each 5. Column Purification, elute with 40 uL EB 6. Measure concentrations in nanodrop