EricChu:LabNotesMDA/2013-9-15

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MDA Device v9 (PGP1 #16) with Tagmentation[edit]

  • Device preparation was described in EricChu:LabNotesMDA/2013-5-20
  • MDA recipe was followed to the protocol in EricChu:LabNotesMDA/2013-5-14
  • Compare experiments of (1) v7 device with Tagmentation and (2) v9 device with Tagmentation
  • One PGP1 cell was used in this experiment.

ALS and NS in MDA Device v9[edit]

  • 240mM KOH final mixing concentration was used to lyze cell and denature DNA

ALS[edit]

H2O 76.78 uL
2M DTT 5 uL
5M KOH 5.72 uL
8% Tween20 12.5 uL

NS[edit]

H2O 24.94 uL
NS stock 62.56 uL
8% Tween20 12.5 uL

MDA Master Mix Recipe for v9[edit]

Phi29 10x buffer 2.33 uL
200uM N6 primer 8.45 uL
25mM dNTP 1.688 uL
8% Tween20 2.33 uL
Phi29 (1ug/ul, High Conc. Epicentre) 0.84 uL
H2O 4.53 uL
  • Preparing "negative" control: added 0.35ul of water into 10ul MDA mastermix in PCR tube. 15 hours at 30C.

Experimental Comments on Device v9[edit]

  1. Only the first chamber was not completely filled in this device.
  2. The mixing chamber was not amplified.