Team:Paris Saclay/Notebook/September/3

From 2014.igem.org

(Difference between revisions)
(→PCR LS)
(→LabWork)
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[photo] Electrophoresis after purification
[photo] Electrophoresis after purification
-
====Ligation===
+
====Ligation====
We already have some pPSI digested and dephosphorelated
We already have some pPSI digested and dephosphorelated
So I do a ligation:
So I do a ligation:
 +
{| class="wikitable centre" width="50%"
 +
|+
 +
|-
 +
! scope=col | component
 +
! scope=col | volume
 +
|-
 +
|H<sub>2</sub>O
 +
|5μl
 +
|-
 +
|buffer
 +
|2μl
 +
|-
 +
|ligase
 +
|1μl
 +
|-
 +
|pPSI
 +
|2μl
 +
|-
 +
|LS PCR
 +
|10μl
 +
|}
 +
 +
2 hours at room temperature and over night at 4°
 +
 +
====pPS5====
 +
Plasmid exctraction using the kit
 +
digestion by SalI
 +
{| class="wikitable centre" width="50%"
 +
|+
 +
|-
 +
! scope=col | component
 +
! scope=col | volume
 +
|-
 +
|H<sub>2</sub>O
 +
|11μl
 +
|-
 +
|buffer
 +
|5μl
 +
|-
 +
|SalI
 +
|2μl
 +
|-
 +
|pPS5
 +
|30μl
 +
|}
 +
Electrophoresis
 +
 +
[photo]
 +
 +
We saw that we have something strang with our plasmid
 +
 +
===pPS3 and pPS4===

Revision as of 15:53, 4 September 2014

Contents

LabWork

B- Construction of the fusion protein

We made a classic exttraction of plasmids from the liquid cultures.

Check by electrophoresis if it worked and send it for sequencing.


D- Lemon scent

by Mélanie

PCR LS

Electrophoresis of the PCR made Yesterday

[photo]

The PCR have success so I use the PCR clean up kit to purify it

[photo] Electrophoresis after purification

Ligation

We already have some pPSI digested and dephosphorelated

So I do a ligation:

component volume
H2O 5μl
buffer 2μl
ligase 1μl
pPSI 2μl
LS PCR 10μl

2 hours at room temperature and over night at 4°

pPS5

Plasmid exctraction using the kit digestion by SalI

component volume
H2O 11μl
buffer 5μl
SalI 2μl
pPS5 30μl

Electrophoresis

[photo]

We saw that we have something strang with our plasmid

pPS3 and pPS4

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