EricChu:LabNotesMDA/2013-9-10: Difference between revisions

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== MDA Device v7 (PGP1 #?) with Tagmentation ==
== MDA Device v7 (PGP1 #15) with Tagmentation ==


* Device preparation was described in EricChu:LabNotesMDA/2013-5-20
* Device preparation was described in EricChu:LabNotesMDA/2013-5-20
* 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 experiments of (1) v7 device with Tagmentation and (2) v9 device 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
|}
* Preparing "negative" control: added 0.35ul of water into 10ul MDA mastermix in PCR tube. 15 hours at 30C.
 
====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

Latest revision as of 01:09, 16 September 2013

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