EricChu:LabNotesMDA/2013-9-15

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Revision as of 04:13, 16 September 2013 by >Ericchu
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MDA Device v9 (PGP1 #16) with Tagmentation

  • 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

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

ALS

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

NS

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

MDA Master Mix Recipe for v9

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.

Comments on Device v9

  1. Only the first chamber was not completely filled in this device.