Daniel:Notebook/HiResChrPaint/2014-1-8
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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