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Team:Paris Saclay/Notebook/August/8
From 2014.igem.org
(→D - Lemon Scent) |
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==Lab work== | ==Lab work== | ||
===D - Lemon Scent=== | ===D - Lemon Scent=== | ||
| - | + | ====PCR==== | |
''by Mélanie & Pierre'' | ''by Mélanie & Pierre'' | ||
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We have done a PCR with pcola (geranyl synthase) and BBa_K517003: | We have done a PCR with pcola (geranyl synthase) and BBa_K517003: | ||
| - | Green GotaqBuffer = 10µl | + | * Green GotaqBuffer = 10µl |
| + | * MgCl2 = 4µl | ||
| + | * dNTP = 1µl | ||
| + | * Primer A = 1µM | ||
| + | * Primer B = 1µM | ||
| + | * DNA = 1µl | ||
| + | * Gotaq DNA polymerase = 0.25µl | ||
| - | + | Oligo used : iPS81bis AND iPS82 (pcola) iPS69 and iPS68bis (BBa_K517003) | |
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| - | Oligo used : iPS81bis AND iPS82 (pcola) | + | |
PCR cycles: | PCR cycles: | ||
| - | 95° 2min | + | * 95° 2min |
| - | + | * 95° 30sec | |
| - | 95° 30sec | + | * 49-55° 30 sec |
| - | + | * 72° 2min | |
| - | 49-55° 30 sec | + | * 71° 5min |
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| - | 72° 2min | + | |
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| - | 71° 5min | + | |
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| + | ====Electrophoresis==== | ||
| + | [[File:Paris Saclay-080814-Pierre-Mélanie-PCR.jpg|right|400px]] | ||
#10 µl of '''BBa_K517003''' | #10 µl of '''BBa_K517003''' | ||
#10 µl of '''BBa_K517003''' | #10 µl of '''BBa_K517003''' | ||
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#10 µl of '''p. cola''' | #10 µl of '''p. cola''' | ||
| - | + | ====Purification of the previous PCR==== | |
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| - | Purification of the previous PCR | + | |
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''by Pierre'' | ''by Pierre'' | ||
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#1 µl of '''p. cola''' | #1 µl of '''p. cola''' | ||
| - | + | ====Stock of pcola plasmides==== | |
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| - | ==== | + | |
using the DNA kit extraction to extract pcola () | using the DNA kit extraction to extract pcola () | ||
Revision as of 10:12, 11 August 2014
Contents |
Friday 8th August
Lab work
D - Lemon Scent
PCR
by Mélanie & Pierre We have done a PCR with pcola (geranyl synthase) and BBa_K517003:
- Green GotaqBuffer = 10µl
- MgCl2 = 4µl
- dNTP = 1µl
- Primer A = 1µM
- Primer B = 1µM
- DNA = 1µl
- Gotaq DNA polymerase = 0.25µl
Oligo used : iPS81bis AND iPS82 (pcola) iPS69 and iPS68bis (BBa_K517003)
PCR cycles:
- 95° 2min
- 95° 30sec
- 49-55° 30 sec
- 72° 2min
- 71° 5min
Electrophoresis
- 10 µl of BBa_K517003
- 10 µl of BBa_K517003
- 10 µl of BBa_K517003
- 10 µl of BBa_K517003
- 10 µl of BBa_K517003
- 10 µl of p. cola
- 10 µl of p. cola
- 10 µl of p. cola
- 10 µl of p. cola
- 10 µl of p. cola
Purification of the previous PCR
by Pierre
From the 5 tubes of p.Cola I cleaned up two tubes. From the 5 tubes of BBa_K517003 I cleaned up two tubes.
I did an electroforesis to check if the clening has worked. It did.
- 1 µl of BBa_K517003
- 1 µl of p. cola
Stock of pcola plasmides
using the DNA kit extraction to extract pcola ()
Human Pratices
Art & Design
by Terry
Sequel of yesterday molding attempt.
The mold is cut delicately and the test piece removed.
Then, the mold is placed back in the becher and filled up with regular agar ( 20mg/ml )
Result : FAILURE, the agar test piece has been ripped when unmolding. It was stuck to the mold.
But the mold is still intact ! I cleaned it up and slightly covered the inward surface with liquid soap. The mold is filled up again and conserved in the refrigerator for 2 hours.
Result : SUCCESS, I slightly remoistened the gel with water for an easier unmolding. The agar test piece is perfectly removed.
