Daniel:Notebook/HiResChrPaint/2014-1-8

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Revision as of 22:22, 8 January 2014 by >Djacobse (→‎Nanodrop Results)
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Alu Control

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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

qPCR Results

File:2014-01-08-Alu1.png

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