Team:Paris/September 8

From 2008.igem.org

(Difference between revisions)
(condition used)
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|400 U
|400 U
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 +
==Digestion==
 +
reaction mixture
 +
(carried  out four times )
 +
* ADN (MP101.1)  3.8 µL
 +
* H2O 21.9 µL
 +
* Buffer10x 3 µL
 +
* BSA 0.3 µl
 +
* EcoRI 1 µL

Revision as of 14:21, 9 September 2008

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Checking our ligases

Since our ligation experiments didn't work during these few days, we presume that our problem came from the ligases. That's why we decided to check the activity of the ligases in our possession. To do so:

  • we will digest a plasmid with only one restriction enzyme (EcoRI).
  • then we will try to ligate it back using our ligases.

condition tested

ligase incubation time amount of enzyme
L1 (ligase 1) 2h00 2 U (unit)
L1 2h00 400 U
L1 O/N 2 U
L1 O/N 400 U
L2 (ligase 2) 2h00 2 U
L2 2h00 400 U
L2 O/N 2 U
L2 O/N 400 U

Digestion

reaction mixture (carried out four times )

  • ADN (MP101.1) 3.8 µL
  • H2O 21.9 µL
  • Buffer10x 3 µL
  • BSA 0.3 µl
  • EcoRI 1 µL