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Team:Evry/Notebook/w8
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<h2>Friday, 3nd August</h2> | <h2>Friday, 3nd August</h2> | ||
| - | <h3> | + | <h3>Construction pCS2+ biobricked:</h3> |
| - | PCR pCS2+ | + | PCR pCS2+ Biobricked |
Result gel:<br/> | Result gel:<br/> | ||
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<img src="https://static.igem.org/mediawiki/2012/1/18/Capture_01423.JPG" alt="A gel width="500" height="500" " /> | <img src="https://static.igem.org/mediawiki/2012/1/18/Capture_01423.JPG" alt="A gel width="500" height="500" " /> | ||
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| + | 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.<br/> | ||
| + | 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. | ||
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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:
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.
