Daniel:Notebook/HiResChrPaint/2013-7-30: Difference between revisions

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>Djacobse
(Created page with "=S1 Probe Preparation= Making a new batch of S1 probe, since I have the other three. ==PCR Amplification== 1. Reaction Table {| class="wikitable" <hiddentext>generated wi...")
 
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|- style="font-size:12pt" align="center"
|- style="font-size:12pt" align="center"
| height="30" | V4S2 (12.5x)
| height="30" | V4S1 (12.5x)
| align="center" | 2.5
| align="center" | 2.5
| align="center" | 625
| align="center" | 625
Line 45: Line 45:


|- style="font-size:12pt" align="center"
|- style="font-size:12pt" align="center"
| height="30" | V6S2 (12.5x)
| height="30" | V6S1 (12.5x)
| align="center" | 2.5
| align="center" | 2.5
| align="center" | 625
| align="center" | 625

Revision as of 21:04, 30 July 2013

S1 Probe Preparation

Making a new batch of S1 probe, since I have the other three.

PCR Amplification

1. Reaction Table
  20 nM OligoPool 2X Kapa SYBR qPCR MM 100 uM AP1V41U 100 uM AP2V4 100 uM AP1V61U 100 uM AP2V6 2 mM aa-dUTP H20 Total Volume
per rxn 0.2 50 0.4 0.4 0.4 0.4 2.5 45.7 100
V4S1 (12.5x) 2.5 625 5 5 0 0 31 581.5 1250
V6S1 (12.5x) 2.5 625 0 0 5 5 31 581.5 1250
2. Aliquot 100 uL into each of 12 qPCR tubes 
3. PCR protocol:
    i. PCR 3 min 95C
    ii. 95C 3 sec
    iii. 55C 30 sec 
    iv. 72C 20 sec
    v. plate read
    vi. goto ii x17
    vii. 72C 2 min
    viii. 16C hold
4. Pooled 3 samples together each (300 uL each, 4 pools per set, 8 total)
5. Column Purification, elute with 40 uL EB
6. Measure concentrations in nanodrop

qPCR Results

Nanodrop Results