EricChu:LabNotesMDA/2014-1-22

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MDA in v9 for testing genomic DNA with Low Concentration KOH denaturation[edit]

  • ~6pg in a single cell
  • 0.3nl volume per compartment and 24 compartments = 7.2nl volume
  • single cell DNA concentration = 6pg/7.2nl = 0.833ng/ul
  • Prepared 40mM KOH and mixed with 1.66ng/ul human genomic DNA in 1:1 ratio. Final KOH concentration = 20mM.
  • NO BSA/N6 primers coating of the MDA chambers in this experiment.

recipe[edit]

0.8ul 5M KOH 5ul 2M DTT 0.1ul 0.5M EDTA 12.5ul 8% Tween20 81.6ul water

result[edit]

  • video was taken of the above experiment
  • DNA was seen after 15hr at 30C and collected in 5ul TE.
  • 1ul sample from each chamber was used to run qPCR with Alu repeat primers.
  • all samples were amplified ok in the device. BSA coating may not be needed.

File:01232014qPCRAluMDA.jpg