Team:Paris Liliane Bettencourt/Notebook/2011/08/29/

From 2011.igem.org

(Difference between revisions)
(→Ligation)
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* 50ng of vector DNA
* 50ng of vector DNA
-
*** m<sub>insert</sub> = Ratio X m<sub>vector</sub> x BP<sub>insert</sub> : BP<sub>vector</sub>
+
* m<sub>insert</sub> = Ratio X m<sub>vector</sub> x BP<sub>insert</sub> : BP<sub>vector</sub>
 +
 
 +
That gives:
 +
 
 +
For the ratio 1:1: 21 ng of insert
 +
For the ratio 1:6: 127 ng of insert
 +
 
 +
 
 +
The YFP-TetR will be ligated on itself. This manipulation removes the additional PstI site.
 +
 
 +
Only the tube one will be carried on.

Revision as of 12:53, 29 August 2011

Team IGEM Paris 2011

Cyrille

Digestion

New attempt of ligation and destroying the next PstI site.

2 times 500ng of S24 was digested in SP for 3h 2 times 500ng of QCP YFP TetR was digested in P for 3h 500ng from the tubes 1 and 6 was digested in XP for 1h and the digestion runned on a gel.

A gel extraction of the band was carried on the lione 1. The result is about 9ng/µL.

ladder - 1 - 6
Cutted band

Ligation

The S24 and YFP will be ligagted together.

A ratio of 1:1 and 1:6 will be explored.

  • S24: 3,3 kb
  • YFP-TetR: 1,4 kb
  • 50ng of vector DNA
  • minsert = Ratio X mvector x BPinsert : BPvector

That gives:

For the ratio 1:1: 21 ng of insert For the ratio 1:6: 127 ng of insert


The YFP-TetR will be ligated on itself. This manipulation removes the additional PstI site.

Only the tube one will be carried on.

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