Daniel:Notebook/HiResChrPaint/2014-1-8: Difference between revisions
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|style="background-color:#D9D9D9;font-weight:bold" width="65" | gDNA | |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" | 2X Kapa SYBR qPCR MM | ||
|style="background-color:#D9D9D9;font-weight:bold" width="65" | 100 uM | |style="background-color:#D9D9D9;font-weight:bold" width="65" | 100 uM AP1Alu | ||
|style="background-color:#D9D9D9;font-weight:bold" width="65" | 100 uM | |style="background-color:#D9D9D9;font-weight:bold" width="65" | 100 uM AP2Alu | ||
|style="background-color:#D9D9D9;font-weight:bold" width="65" | H20 | |style="background-color:#D9D9D9;font-weight:bold" width="65" | H20 | ||
|style="background-color:#D9D9D9;font-weight:bold" width="65" | Total Volume | |style="background-color:#D9D9D9;font-weight:bold" width="65" | Total Volume |
Revision as of 22:23, 8 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 AP1Alu | 100 uM AP2Alu | 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
qPCR Results
Nanodrop Results
Sample | ALU1-A | ALU1-B | ALU1-C | ALU1-D |
ng/uL | 15.4 | 13.5 | 15 | 15.9 |
ug in 40 uL | 0.6 | 0.5 | 0.6 | 0.6 |
total | 2.4 |