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