Rui:LabNotes/Haplotyping/2011-11-3
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Test condition for haplotyping[edit]
- Start with ~1pg cell lysate aliquot
- Perform limited time MDA
- Tagmentation and PCR
- Need to achieve ~1000X increase. Based on 50cell experiments, it seems 1k-2k increase after 2hrs when QPCR shoots up around 12ct and reach the saturation at 20ct.
- Key point is when to stop MDA based on QPCR curve
Samples[edit]
Procedures[edit]
Results[edit]
Ctrls File:11.3.11 Ctrls.jpg
@17ct, #2 up File:11.3.11 17ct.jpg @22ct, #3 up File:11.3.11 22ct.jpg
@27ct, #3,4 start to up File:11.3.11 27ct.jpg @30ct, all start up File:11.3.11 30ct.jpg
EtOH precipitation[edit]
- Samples: 17-2, 22-3, 27-3,4, 30-1,2,3,4
' | ng/ul | total (ng) | up Ct | stop Ct | dCt | dFlu. |
17-2 | 6.95 | 34.75 | 12 | 17 | 5 | 400 |
22-3 | 15.1 | 75.5 | 12 | 22 | 10 | 500 |
27-3 | 1.29 | 6.45 | 22 | 26 | 4 | 100 |
27-4 | 1.17 | 5.85 | 22 | 26 | 4 | 100 |
30-1 | 21.5 | 107.5 | 20 | 30 | 10 | 300 |
30-2 | 10.1 | 50.5 | 24 | 30 | 6 | 300 |
30-3 | 15.8 | 79 | 20 | 30 | 10 | 400 |
30-4 | 6.35 | 31.75 | 27 | 30 | 3 | 100 |