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

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=== Ligation ===
=== 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
 +
 +
* 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.
 +
 +
 +
==Axel==
 +
===knock out of CodY gene of the strain php13-v10-6b===
 +
nothing on the plates
 +
We should re-do the extraction of genomic DNA
 +
 +
 +
==Camille==
 +
 +
Every of my strains grew. I selected three clons of each and tested them with a PCR colony. Only two clons seemed to be good for the transformation of the ligation of the tRNA alone with the promoter before.
 +
<br><br><br><br><br>

Latest revision as of 15:15, 3 September 2011

Team IGEM Paris 2011

Contents

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.


Axel

knock out of CodY gene of the strain php13-v10-6b

nothing on the plates We should re-do the extraction of genomic DNA


Camille

Every of my strains grew. I selected three clons of each and tested them with a PCR colony. Only two clons seemed to be good for the transformation of the ligation of the tRNA alone with the promoter before.




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