EricChu:LabNotesMDA/2013-9-15: Difference between revisions
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|12.5 uL | |12.5 uL | ||
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====MDA Master Mix Recipe for v9==== | |||
* The recipe v9 is identical to v7 since no changes in MDA part of the design | |||
*[[EricChu:LabNotesMDA/2013-5-14|EricChu:LabNotesMDA/2013-5-14 protocol]] | |||
{| {{table}} | |||
|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. |
Revision as of 01:34, 16 September 2013
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
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
- 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.