Team:Paris Saclay/Notebook/August/21

From 2014.igem.org

(Difference between revisions)
(→Electro elution of pPS1 digested by Pac1)
(→Lab work)
Line 12: Line 12:
We extracted the DNA from the agarose band saves yesterday  
We extracted the DNA from the agarose band saves yesterday  
 +
 +
 +
==== Ligation of CAD in pMCS5 ====
 +
''by Eugene''
 +
 +
We have made an electrophoresis of purified CAD and pMCS5.
 +
 +
* Dephosphorylation of pMCS5
 +
{| class="wikitable centre" width="50%"
 +
|+
 +
|-
 +
! scope=col | component
 +
! scope=col | volume
 +
|-
 +
|H<sub>2</sub>O
 +
|3 μl
 +
|-
 +
| pMCS5p
 +
| 5 μl
 +
|-
 +
| 10x buffer
 +
| 10 μl
 +
|-
 +
| Fast AP
 +
| 1 μl
 +
|}
 +
 +
30 min incubation in 37°c
 +
 +
* 15min of inactivation 65°c
 +
 +
* Ligation
 +
{| class="wikitable centre" width="50%"
 +
|+
 +
|-
 +
! scope=col | component
 +
! scope=col | volume
 +
|-
 +
|CADp
 +
|10 μl
 +
|-
 +
| pMCS5p
 +
| 2.5 μl
 +
|-
 +
| 10x buffer
 +
| 1.5 μl
 +
|-
 +
| Ligase
 +
| 1 μl
 +
|}
 +
 +
night at 16°c
 +
[https://2014.igem.org/Team:Paris_Saclay/Protocols/Electro_extraction_of_DNA_from_agarose_gel protocol]
[https://2014.igem.org/Team:Paris_Saclay/Protocols/Electro_extraction_of_DNA_from_agarose_gel protocol]
{{Team:Paris_Saclay/notebook_footer}}
{{Team:Paris_Saclay/notebook_footer}}

Revision as of 17:14, 21 August 2014

Contents

Thursday 21st August

Lab work

D - Lemon Scent

Transformation of DH5α with CAD and chromoprotein

by Sean

protocol

Electro elution of pPS1 digested by Pac1

"by Huang vu and laetitia"

We extracted the DNA from the agarose band saves yesterday


Ligation of CAD in pMCS5

by Eugene

We have made an electrophoresis of purified CAD and pMCS5.

  • Dephosphorylation of pMCS5
component volume
H2O 3 μl
pMCS5p 5 μl
10x buffer 10 μl
Fast AP 1 μl

30 min incubation in 37°c

  • 15min of inactivation 65°c
  • Ligation
component volume
CADp 10 μl
pMCS5p 2.5 μl
10x buffer 1.5 μl
Ligase 1 μl

night at 16°c


protocol

Back to the calendar

Retrieved from "http://2014.igem.org/Team:Paris_Saclay/Notebook/August/21"