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]
- The recipe v9 is identical to v7 since no changes in MDA part of the design
- EricChu:LabNotesMDA/2013-5-14 protocol
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]
- Only the first chamber was not completely filled in this device.
- The mixing chamber was not amplified.