Team:Paris Saclay/Notebook/August/26

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(→D - Lemon Scent)
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=Tuesday 26th August=
=Tuesday 26th August=
==Lab work==
==Lab work==
 +
 +
====plasmid extraction : pGEMTeasy + chromoprotein ====
 +
''by Laetitia''
 +
 +
We used the bacteria containing pGEMTeasy + chromoprotein launched by Laetitia monday the 25th for the plasmid extraction, 6 independent cultures which come from the 6 stocks made yesterday.
 +
 +
To extract the plasmid, we used the plasmid DNA purification kit (Macherey-Nagel). Protocol for low copy plasmid.
 +
 +
====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
 +
|10 μl
 +
|-
 +
|Fast Digest buffer 10X
 +
|1,5 μl
 +
|-
 +
|XbaI
 +
|0.5μl
 +
|-
 +
|PstI
 +
|0.5μl
 +
|-
 +
|H20
 +
|2,5 μl
 +
|}
 +
 +
1h at 37°C
 +
 +
==== Electrophoresis of the digestion product of pGEMTeasy+chromoprotein====
 +
''by Laetitia''
 +
 +
T : control negatif without enzyme
 +
 +
{| class="wikitable centre" width="50%"
 +
|+
 +
|-
 +
! scope=col | component
 +
! scope=col | volume
 +
|-
 +
|blue
 +
|2 μl
 +
|-
 +
|Plasmid
 +
|2 μl
 +
|-
 +
|H20
 +
|8μl
 +
|}
 +
 +
 +
D : sample digested by XbaI and PstI
 +
 +
D1 - D2 - D3 - D4 - D5 - D6 - T - L
 +
 +
===D - Lemon Scent===
===D - Lemon Scent===
 +
==Streaking of colonies containing BBa_K762100+pGEMTeasy==
==Streaking of colonies containing BBa_K762100+pGEMTeasy==

Revision as of 14:03, 26 August 2014

Contents

Tuesday 26th August

Lab work

plasmid extraction : pGEMTeasy + chromoprotein

by Laetitia

We used the bacteria containing pGEMTeasy + chromoprotein launched by Laetitia monday the 25th for the plasmid extraction, 6 independent cultures which come from the 6 stocks made yesterday.

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

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 10 μl
Fast Digest buffer 10X 1,5 μl
XbaI 0.5μl
PstI 0.5μl
H20 2,5 μl

1h at 37°C

Electrophoresis of the digestion product of pGEMTeasy+chromoprotein

by Laetitia

T : control negatif without enzyme

component volume
blue 2 μl
Plasmid 2 μl
H20 8μl


D : sample digested by XbaI and PstI

D1 - D2 - D3 - D4 - D5 - D6 - T - L


D - Lemon Scent

Streaking of colonies containing BBa_K762100+pGEMTeasy

Due to yesterday's strange results, we conduct 30 streakings in a petri dish: 28 white colonies and 2 blue colonies.


Back to the calendar

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