Team:Paris Saclay/digestion ligation

From 2013.igem.org

(Difference between revisions)
(Created page with "{{Team:Paris_Saclay/incl_debut_generique}} = '''Digestion and Ligation''' = {{Team:Paris_Saclay/incl_fin}}")
(→Digestion and Ligation)
Line 2: Line 2:
= '''Digestion and Ligation''' =
= '''Digestion and Ligation''' =
 +
 +
Digestion :
 +
 +
{| class="wikitable centre" width="80%"
 +
|+
 +
|-
 +
! scope=col | Enzyme
 +
! scope=col | EcoRI
 +
! scope=col | PstI
 +
! scope=col | SpeI
 +
! scope=col | XbaI
 +
|-
 +
| width="33%" |
 +
Tampon
 +
| width="34%" |
 +
Fast Digest
 +
| width="33%" |
 +
Fast Digest
 +
| width="34%" |
 +
Fast Digest
 +
| width="33%" |
 +
Fast Digest
 +
|-
 +
| width="33%" |
 +
site
 +
| width="34%" |
 +
5'-G/AATTC-3'
 +
3'-CTTAA/G-5'
 +
| width="33%" |
 +
5'CTGCA/G 3'
 +
3'G/ACGTC 5'
 +
| width="34%" |
 +
5'A/CTAGT 3'
 +
3'T G ATC/A5'
 +
| width="33%" |
 +
5'T/CTAGA3'
 +
3'AGATC/T5'
 +
 +
 +
 +
|}
 +
 +
 +
 +
Double Digestion :
 +
{| class="wikitable centre" width="80%"
 +
|+
 +
|-
 +
! scope=col | DNA
 +
! scope=col | volume
 +
|-
 +
| width="33%" |
 +
Tampon FD 10X     
 +
| width="34%" |
 +
3µL
 +
|-
 +
| width="33%" |
 +
Enzyme 1             
 +
| width="34%" |
 +
1.5 μl
 +
|-
 +
| width="33%" |
 +
Enzyme 2
 +
| width="34%" |
 +
1.5 μl
 +
|-
 +
| width="33%" |
 +
h20
 +
| width="34%" |
 +
14µL
 +
 +
 +
 +
 +
|}
 +
 +
 +
Simple Digestion :
 +
{| class="wikitable centre" width="80%"
 +
|+
 +
|-
 +
! scope=col | DNA
 +
! scope=col | volume
 +
|-
 +
| width="33%" |
 +
Tampon FD 10X     
 +
| width="34%" |
 +
3µL
 +
|-
 +
| width="33%" |
 +
Enzyme             
 +
| width="34%" |
 +
1.5 μl
 +
|-
 +
| width="33%" |
 +
h20
 +
| width="34%" |
 +
15.5µL
 +
 +
 +
 +
 +
|}
 +
 +
 +
1 .Incubate at 37°C during 10-90min
 +
2. Inactivate digestion enzyms with ethanol precipitation
 +
 +
Ligation:
 +
Mix in a molar ratio the insert and the linear vector

Revision as of 20:49, 23 September 2013

Navigation Home Team Project Labwork Notebook Human practices

Digestion and Ligation

Digestion :

Enzyme EcoRI PstI SpeI XbaI

Tampon

Fast Digest

Fast Digest

Fast Digest

Fast Digest

site

5'-G/AATTC-3' 3'-CTTAA/G-5'

5'CTGCA/G 3' 3'G/ACGTC 5'

5'A/CTAGT 3' 3'T G ATC/A5'

5'T/CTAGA3' 3'AGATC/T5'



Double Digestion :

DNA volume

Tampon FD 10X

3µL

Enzyme 1

1.5 μl

Enzyme 2

1.5 μl

h20

14µL




Simple Digestion :

DNA volume

Tampon FD 10X

3µL

Enzyme

1.5 μl

h20

15.5µL




1 .Incubate at 37°C during 10-90min 2. Inactivate digestion enzyms with ethanol precipitation

Ligation: Mix in a molar ratio the insert and the linear vector




Retrieved from "http://2013.igem.org/Team:Paris_Saclay/digestion_ligation"