Rui:LabNotes/Haplotyping/2011-11-3

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Revision as of 01:19, 4 November 2011 by >RuiLiu (Created page with '==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 expe…')
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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

stop at 17ct, #2 up File:11.3.11 17ct.jpg stop at 22ct, #3 up File:11.3.11 22ct.jpg

stop at 27ct, #3,4 start to up File:11.3.11 27ct.jpg stop at 30ct, all start up File:11.3.11 30ct.jpg