Team:Paris Saclay/Notebook/September/3

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===pPS3 and pPS4===
===pPS3 and pPS4===
 +
We digest the plasmid by HindIII to direct our insert
 +
 +
! scope=col | component
 +
! scope=col | volume
 +
|-
 +
|H<sub>2</sub>O
 +
|6μl
 +
|-
 +
|buffer
 +
|1μl
 +
|-
 +
|HindIII
 +
|1μl
 +
|-
 +
|pPS3/4
 +
|2μl
 +
|}
 +
 +
[photo]
 +
 +
results are very strange

Revision as of 16:24, 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

We digest the plasmid by HindIII to direct our insert

! scope=col | component ! scope=col | volume |- |H2O |6μl |- |buffer |1μl |- |HindIII |1μl |- |pPS3/4 |2μl |}

[photo]

results are very strange

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