Team:Paris/10 August 2009
From 2009.igem.org
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===Lab work=== | ===Lab work=== | ||
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<div class="guillaume"> | <div class="guillaume"> | ||
PCR | PCR | ||
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<div class="experience"> | <div class="experience"> | ||
- | *For [https://2009.igem.org/Team:Paris/Freezer_PCR_products#A8 A8] and [https://2009.igem.org/Team:Paris/Freezer_PCR_products#A9 A9] | + | *For [https://2009.igem.org/Team:Paris/Freezer_PCR_products#A8 A8]OmpA-Linker and [https://2009.igem.org/Team:Paris/Freezer_PCR_products#A9 A9]pFecA |
+ | *Duplicate with and without DMSO | ||
**1:98°C 1min | **1:98°C 1min | ||
- | **2:98°C 10s | + | **2:98°C 10s |
- | **3:59°C | + | **3:59°C for A8) / 65°C for A9 30s |
**4:72°C 8s | **4:72°C 8s | ||
**5:72°C 10min | **5:72°C 10min | ||
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|width=10%| D0(320) | |width=10%| D0(320) | ||
|width=10%| Dilution Rule | |width=10%| Dilution Rule | ||
+ | |width=10%| Primer Addition | ||
|width=10%| Water Addition | |width=10%| Water Addition | ||
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|width=10%| 0.004 | |width=10%| 0.004 | ||
|width=10%| 600/310 = 1.93 ~ 2.0 µL | |width=10%| 600/310 = 1.93 ~ 2.0 µL | ||
- | |width=10%| 2.45 | + | |width=10%| 2.45 µL |
+ | |width=10%| 10.55 µL | ||
|- style="background: #E0E9EF; text-align: center;" | |- style="background: #E0E9EF; text-align: center;" | ||
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|width=10%| 0.000 | |width=10%| 0.000 | ||
|width=10%| 600/134 = 4.48 (~ 4.5 µL) | |width=10%| 600/134 = 4.48 (~ 4.5 µL) | ||
- | |width=10%| 2.45 | + | |width=10%| 2.45 µL |
+ | |width=10%| 8.05 µL | ||
|- style="background: #E0E9EF; text-align: center;" | |- style="background: #E0E9EF; text-align: center;" | ||
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|width=10%| 0.000 | |width=10%| 0.000 | ||
|width=10%| 600/70 = 8.57 ~ 8.6 µL | |width=10%| 600/70 = 8.57 ~ 8.6 µL | ||
- | |width=10%| 2. | + | |width=10%| 2.45µL |
- | + | |width=10%| 3.95µL | |
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<div class="experience"> | <div class="experience"> | ||
- | + | GEL!!! | |
- | + | Remigrate to be sure... pFecA seem to be good but not Ompa-Linker | |
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Revision as of 23:40, 10 August 2009
Contents |
NoteBook
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Brain work
edit please ^^
Lab work
PCR
Mini-Prep for Plac
Same Protocol as usual ...
Do
Dilution for Plasmids 1,2&3 | ||||||||||
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Plasmid | D0(000) | D0(266/280) | D0(320) | Dilution Rule | Primer Addition | Water Addition | ||||
P1 | 3.10 | 1.72 | 0.004 | 600/310 = 1.93 ~ 2.0 µL | 2.45 µL | 10.55 µL | ||||
P2 | 1.34 | 1.34 | 0.000 | 600/134 = 4.48 (~ 4.5 µL) | 2.45 µL | 8.05 µL | ||||
P3 | 0.70 | 3.39 | 0.000 | 600/70 = 8.57 ~ 8.6 µL | 2.45µL | 3.95µL |
Gel Migration
GEL!!! Remigrate to be sure... pFecA seem to be good but not Ompa-Linker
Purification
purification with promega kits gel purification -> Very good
To do list
Matricule | TODO |
Luc | |
Romain | |
Charlotte | |
Stoff | |
Chris | |
Lisa | |
Caroline | |
Souf | |
Vicard | |
Pierre | |
Sylvain | |
Guillaume |