EricChu:LabNotesMDA/2013-9-15: Difference between revisions

From ZhangLabWiki
Jump to navigation Jump to search
>Ericchu
No edit summary
>Ericchu
No edit summary
Line 41: Line 41:
|12.5 uL
|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|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

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.