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" | &nbsp;
|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

Back to Calendar

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