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Team:Paris Saclay/Notebook/September/4
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| - | {{Team:Paris_Saclay/ | + | {{Team:Paris_Saclay/notebook_header}} |
| + | =Thursday 4st September= | ||
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==Lab Work== | ==Lab Work== | ||
''By Mélanie'' | ''By Mélanie'' | ||
| - | === | + | ===Lemon Scent=== |
====Verfication of the pPS3/4/5 plasmid==== | ====Verfication of the pPS3/4/5 plasmid==== | ||
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I use the same PCR condition than [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/2#Limonene_synthase September 2] but with the right primer and plasmid | I use the same PCR condition than [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/2#Limonene_synthase September 2] but with the right primer and plasmid | ||
| - | [[File:0309 Vérif PCR GS CAD PS LS Chromo.jpg|400px]] | + | [[File:0309 Vérif PCR GS CAD PS LS Chromo.jpg|400px|center]] |
We can see that we don't have any insert in our plasmid | We can see that we don't have any insert in our plasmid | ||
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We find some other PCR of BBa K762100 and GS and BBa_K517003 so we check it by electrophoresis | We find some other PCR of BBa K762100 and GS and BBa_K517003 so we check it by electrophoresis | ||
| - | [[File:0309 Vérif PCR GS CAD PS LS Chromo.jpg|400px]] | + | [[File:0309 Vérif PCR GS CAD PS LS Chromo.jpg|400px|center]] |
And I purify the PCR | And I purify the PCR | ||
| - | [[File:0309 PCR cleanup GS PS CAD.jpg|400px]] | + | [[File:0309 PCR cleanup GS PS CAD.jpg|400px|center]] |
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Transformation with competent bacteria | Transformation with competent bacteria | ||
| - | add DNA (Top vector | + | add DNA (Top vector) |
30' on ice | 30' on ice | ||
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spread on petri dish | spread on petri dish | ||
| - | {{Team:Paris_Saclay/ | + | |
| + | ==Photo of the Day== | ||
| + | [[File:Paris Saclay 4_september.jpg|600px|center]] | ||
| + | |||
| + | {{Team:Paris_Saclay/notebook_footer}} | ||
Latest revision as of 12:52, 14 October 2014
Contents |
Thursday 4st September
Lab Work
By Mélanie
Lemon Scent
Verfication of the pPS3/4/5 plasmid
Due to the strange results obtained last day, I do a PCR of the insert with the differents plasmid : I use the same PCR condition than September 2 but with the right primer and plasmid
We can see that we don't have any insert in our plasmid
PCR verification
We find some other PCR of BBa K762100 and GS and BBa_K517003 so we check it by electrophoresis
And I purify the PCR
Well 1 = Ladder
Well 2 = GS
Well 3 = PS
Well 4 = CAD
We can see that we don't have lost a lot of our PCR product
cloning
Cloning of PCR purification in a TA plasmid (Topo plasmid)
First I had to add AAA to the PCR
| component | volume |
|---|---|
| buffer | 1μl |
| dATP | 1μl |
| PCR | 7.5μl |
| Taq | 0.5μl |
72° 15'
and
| component | volume |
|---|---|
| H2O | 1μl |
| buffer | 1μl |
| vector | 1μl |
| cloning | 1μl |
30' at room temperature
Transformation
Transformation with competent bacteria
add DNA (Top vector)
30' on ice 45' 42°c 30' at 37°
spread on petri dish
Photo of the Day
