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Team:Paris Saclay/Notebook/September/3
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The PCR have success so I use the PCR clean up kit to purify it | The PCR have success so I use the PCR clean up kit to purify it | ||
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| + | [photo] | ||
====Ligation==== | ====Ligation==== | ||
Revision as of 11:53, 7 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
Electrophoresis after purification
[photo]
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
We saw that we have something strang with our plasmid but we wil check it tomorrow with a good ladder (the ladder here was normally on the well 1)
pPS3 and pPS4
We digest the plasmid by HindIII to direct our insert
| component | volume |
|---|---|
| H2O | 6μl |
| buffer | 1μl |
| HindIII | 1μl |
| pPS3/4 | 2μl |
well 1 = ladder well 2-3-4 = pPS3 Well 5-6-7 = pPS4
results are very strange
