"

From 2012.igem.org

Revision as of 18:17, 3 August 2012 (view source) TriCer (Talk | contribs) ← Older edit		Latest revision as of 18:19, 3 August 2012 (view source) TriCer (Talk | contribs)
Line 50:		Line 50:
	<center>		<center>
	1) is PCR products with template pCS2+, primers P7 and P8    and  2) to 6) are ladders test		1) is PCR products with template pCS2+, primers P7 and P8    and  2) to 6) are ladders test
-		+	<br/><br/>
-	Then the PCR product pCS2+ was purified with kit, digested with EcoRI and PstI then purified with PCR cleanup kit.	+	Then the PCR product pCS2+ was purified with kit, digested with EcoRI and PstI then purified with PCR cleanup kit.<br/>
	Plasmid TSO 28.06 mRFP (plasmid from plate 1-18F, mRFP)was cut with EcoRI and PstI:		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.		after electrophoresis no band visible, 7 microL of plasmid used in a total volume of 25 microL.

---

Latest revision as of 18:19, 3 August 2012

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.