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From 2012.igem.org

Weeks:

June			July				August				September				October				November

Week 8: 30th July - 5th August

Monday, 30th July

Digestion - Ligation - Transformation:

Digestion of psc2+ and mRFP with EcoRI and PstI.
Ligation and Transformation on T10.

TirI BB

Electrophoresis of TirI last PCR: failure of this technique.
We buy a new primer to try another PCR technic.

Tuesday, 31st July

Inoculation:

Inoculation of pcs2+ (27.07) and repiquage of psc2+ with RFP (30.07)
Inoculation of psB1C3 with GFP-AID TirI: Electrophoresis of 27/07 results: Failure of second PCR
We restart the first step

Thursday, 2nd August

Sequencing

Send pCS2+ to sequencing.

Issue

We found that pCS2+ was badly made. We restart the construction of this plasmid from the beginning.

Friday, 3nd August

Construction pCS2+ biobricked:

PCR pCS2+ Biobricked Result gel:
1) is PCR products with template pCS2+, primers P7 and P8 and 2) to 6) are ladders test

Then the PCR product pCS2+ was purified with kit, digested with EcoRI and PstI then purified with PCR cleanup kit.
Plasmid TSO 28.06 mRFP (plasmid from plate 1-18F, mRFP)was cut with EcoRI and PstI: after electrophoresis no band visible, 7 microL of plasmid used in a total volume of 25 microL.