Team:Paris Saclay/Notebook/August/25

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(→D - Lemon Scent)
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=Monday 25th August=
=Monday 25th August=
==Lab work==
==Lab work==
 +
 +
===B - Construction of the fusion protein===
 +
 +
====plasmid extraction : pGEMTeasy + chromoprotein ====
 +
''by Laetitia''
 +
 +
We used the bacteria containing pGEMTeasy + chromoprotein launched by Melanie sunday the 24th for the plasmid extraction, 6 independent cultures which come from 6 independent colonies.
 +
 +
We made a stock of bacteria for each sample (x6).
 +
 +
To extract the plasmid, we used the plasmid DNA purification kit (Macherey-Nagel).
 +
 +
====Digestion of pGEMTeasy+chromoproteinby XbaI and PstI====
 +
''by Laetitia''
 +
 +
The goal is to check the presence of the insert inside the plasmid. We digest the 6 samples of purified plasmid.
 +
 +
{| class="wikitable centre" width="50%"
 +
|+
 +
|-
 +
! scope=col | component
 +
! scope=col | volume
 +
|-
 +
|Plasmid
 +
|5 μl
 +
|-
 +
|Fast Digest buffer 10X
 +
|1μl
 +
|-
 +
|XbaI
 +
|0.5μl
 +
|-
 +
|PstI
 +
|0.5μl
 +
|-
 +
|H20
 +
|3μl
 +
|}
 +
 +
1h at 37°C
 +
 +
==== Electrophoresis of the digestion product of pGEMTeasy+chromoprotein====
 +
''by Laetitia''
 +
 +
(image)
 +
 +
Results : the samples 3, 4 and 6 seems to contain the chromoprotein gene.
 +
 +
==== Bacterial culture of DH5a containing pGEMTeasy + chromprotein ====
 +
''by Laetitia''
 +
 +
We launched 6 cultures in 5mL LB + Ampi (1/1000) - 37°C at night . The bacteria come from the stock made before the plasmid extraction
 +
 +
===D - Lemon Scent===
===D - Lemon Scent===
====Gel electrophoresis of CAD====
====Gel electrophoresis of CAD====

Revision as of 14:37, 25 August 2014

Contents

Monday 25th August

Lab work

B - Construction of the fusion protein

plasmid extraction : pGEMTeasy + chromoprotein

by Laetitia

We used the bacteria containing pGEMTeasy + chromoprotein launched by Melanie sunday the 24th for the plasmid extraction, 6 independent cultures which come from 6 independent colonies.

We made a stock of bacteria for each sample (x6).

To extract the plasmid, we used the plasmid DNA purification kit (Macherey-Nagel).

Digestion of pGEMTeasy+chromoproteinby XbaI and PstI

by Laetitia

The goal is to check the presence of the insert inside the plasmid. We digest the 6 samples of purified plasmid.

component volume
Plasmid 5 μl
Fast Digest buffer 10X 1μl
XbaI 0.5μl
PstI 0.5μl
H20 3μl

1h at 37°C

Electrophoresis of the digestion product of pGEMTeasy+chromoprotein

by Laetitia

(image)

Results : the samples 3, 4 and 6 seems to contain the chromoprotein gene.

Bacterial culture of DH5a containing pGEMTeasy + chromprotein

by Laetitia

We launched 6 cultures in 5mL LB + Ampi (1/1000) - 37°C at night . The bacteria come from the stock made before the plasmid extraction


D - Lemon Scent

Gel electrophoresis of CAD

by Sean

Legend

  1. ladder 10µl
  2. PCR result of CAD

Paris Saclay Sean 140825.jpg

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