EricChu:LabNotesMDA/2013-9-10: Difference between revisions
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* MDA recipe was followed to the protocol in EricChu:LabNotesMDA/2013-5-14 | * MDA recipe was followed to the protocol in EricChu:LabNotesMDA/2013-5-14 | ||
* Compare experiment with Tagmentation | * Compare experiment with Tagmentation | ||
====MDA Master Mix Recipe for v7==== | |||
*[[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 | |||
|} | |||
====ALS, NS buffer==== | |||
'''ALS (240mM after mixing cell capture volume)''' | |||
{| {{table}} | |||
|H2O | |||
|| | |||
|77.12 uL | |||
|- | |||
|2M DTT | |||
|| | |||
|5 uL | |||
|| | |||
|- | |||
|5M KOH | |||
|| | |||
|5.38 uL | |||
|- | |||
|8% Tween20 | |||
|| | |||
|12.5 uL | |||
|} | |||
'''NS (matching ALS)''' | |||
{| {{table}} | |||
|H2O | |||
|| | |||
|28.66 uL | |||
|- | |||
|NS stock | |||
|| | |||
|58.84 uL | |||
|- | |||
|8% Tween20 | |||
|| | |||
|12.5 uL | |||
|} | |||
*NS stock : 4mL of 1M HCl + 6mL of 1M TrisHCl |
Revision as of 22:19, 10 September 2013
MDA Device v7 (PGP1 #?) 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 experiment with Tagmentation
MDA Master Mix Recipe for v7
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 |
ALS, NS buffer
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