Team:Paris Saclay/Notebook/August/27

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==Lab Work==
==Lab Work==
===B - Construction of the fusion protein (color)===
===B - Construction of the fusion protein (color)===
 +
====PCR clean-up of colonies containing pGEMTeasy+chromoprotein====
 +
''by Sean''
 +
 +
Three colonies were selected from the gel prepared earlier by Laetitia, namely #3,#4, and #6. The three gel samples underwent PCR clean-up.
 +
 +
====Gel electrophoresis of colonies containing pGEMTeasy+chromoprotein====
 +
''by Sean''
 +
 +
Gel containing the said colonies underwent PCR clean-up. 1µl of each result was used for the electrophoresis. Nothing appeared under UV.
 +
===D - lemon scent ===
===D - lemon scent ===
====Plasmide extraction====
====Plasmide extraction====

Revision as of 16:49, 27 August 2014

Contents

Wednesday 27th August

Lab Work

B - Construction of the fusion protein (color)

PCR clean-up of colonies containing pGEMTeasy+chromoprotein

by Sean

Three colonies were selected from the gel prepared earlier by Laetitia, namely #3,#4, and #6. The three gel samples underwent PCR clean-up.

Gel electrophoresis of colonies containing pGEMTeasy+chromoprotein

by Sean

Gel containing the said colonies underwent PCR clean-up. 1µl of each result was used for the electrophoresis. Nothing appeared under UV.

D - lemon scent

Plasmide extraction

By Mélanie

Extraction of pPS5 with the phenol protocol (as previously described)

PCR of Limonene synthase (BBa762100)

by Mélanie

5 tubes to do a gradient

components volumes
H2O 29.75μl
Gotaq buffer 10µl
dNTP 10mM 1µl
iPS67 1µM
iPS66 1µM
DNA 1µl
MgCl2 4µl
Gotaq 1.25µl
PCR cycle
step temperature (°C) time
1 95 2min
2 95 30 sec
3 49-55 (gradient) 30sec
4 72 2min
5 72 5min


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