EricChu:LabNotesMDA/2013-9-10
Jump to navigation
Jump to search
MDA Device v7 (PGP1 #15) 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
MDA Master Mix Recipe for v7[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.
ALS, NS buffer[edit]
ALS (240mM after mixing cell capture volume)
H2O | 77.12 uL | ||
2M DTT | 5 uL | ||
5M KOH | 5.38 uL | ||
8% Tween20 | 12.5 uL |
NS (matching ALS)
H2O | 28.66 uL | |
NS stock | 58.84 uL | |
8% Tween20 | 12.5 uL |
- NS stock : 4mL of 1M HCl + 6mL of 1M TrisHCl