Rui:LabNotes/Haplotyping/2011-11-3: Difference between revisions
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===EtOH precipitation=== | ===EtOH precipitation=== | ||
* Samples: 17-2, 22-3, 27-3,4, 30-1,2,3,4 | * Samples: 17-2, 22-3, 27-3,4, 30-1,2,3,4 | ||
{| {{table}} border=1 | |||
| align="center" style="background:#f0f0f0;"|'''''' | |||
| align="center" style="background:#f0f0f0;"|'''ng/ul''' | |||
| align="center" style="background:#f0f0f0;"|'''total (ng)''' | |||
| align="center" style="background:#f0f0f0;"|'''up Ct''' | |||
| align="center" style="background:#f0f0f0;"|'''stop Ct''' | |||
| align="center" style="background:#f0f0f0;"|'''total Ct''' | |||
| align="center" style="background:#f0f0f0;"|'''total Flu.''' | |||
|- | |||
| 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 | |||
|} |
Revision as of 19:06, 4 November 2011
Test condition for haplotyping
- 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
Procedures
Results
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
- Samples: 17-2, 22-3, 27-3,4, 30-1,2,3,4
' | ng/ul | total (ng) | up Ct | stop Ct | total Ct | total Flu. |
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 |