From 2013.igem.org
Contents
- 1 Notebook : July 26
- 1.1 Lab work
- 1.1.1 A - Aerobic/Anaerobic regulation system
- 1.1.1.1 Objective : obtaining Bba_K1155003
- 1.1.1.2 1 - Ligation of RBS-AmilCP and Term in PSB1C3
- 1.1.1.3 Objective : obtaining Bba_K1155004, Bba_K1155005, Bba_K1155006
- 1.1.1.4 1 - Electrophoresis of PCR products : NarK, NarG, NirB
- 1.1.1.5 2 - PCR of NarK, NarG, NirB
- 1.1.1.6 3 - Electrophoresis of PCR products : NarK, NarG, NirB
- 1.1.1.7 4 - Gel purification of PCR products : NarK, NarG, NirB
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Notebook : July 26
Lab work
A - Aerobic/Anaerobic regulation system
Objective : obtaining Bba_K1155003
1 - Ligation of RBS-AmilCP and Term in PSB1C3
Xavier
Used quantities :
- RBS-AmilCP : 2µL
- Bba_B0015 digested by ... : 2.5µL
- Buffer : 1µL
- Ligase : 1µL
- H2O : 3.5µL
Objective : obtaining Bba_K1155004, Bba_K1155005, Bba_K1155006
1 - Electrophoresis of PCR products : NarK, NarG, NirB
Abdou
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- Well 1 : 5µL of NirB+1µL of 6X loading dye
- Well 2 : 5µL of NirB+1µL of 6X loading dye LA MEME CHOSE ???????
- Well 3 : 5µL of NarG+1µL of 6X loading dye
- Well 4 : 5µL of NarG+1µL of 6X loading dye
- Well 5 : 6µL DNA Ladder
- Well 6 : 5µL of NarK+1µL of 6X loading dye NUMERO 4 ???????????????
- Well 7 : 5µL of NarK+1µL of 6X loading dye
- Well 8 : 5µL of NarK+1µL of 6X loading dye
- Gel : 1%
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Expected sizes :
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We lost all our PCR products. We will do PCR again using more concentrated DNA and oligos.
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2 - PCR of NarK, NarG, NirB
Abdou, Xavier
Used quantities : cf PCR précédente !!!!!!!!!!!!!
... ADN non dilué
PCR program : cf PCR précédente !!!!!!!!!
SCHEMA !!!!!!!!!!!!!!
3 - Electrophoresis of PCR products : NarK, NarG, NirB
Abdou, Xavier
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- Well 1 : 5µL of NirB+1µL of 6X loading dye quantité 2µL d'ADN ????????
- Well 2 : 5µL of NirB+1µL of 6X loading dye
- Well 3 : 5µL of NarK+1µL of 6X loading dye
- Well 4 : 5µL of NarK+1µL of 6X loading dye
- Well 5 : 6µL DNA Ladder
- Well 6 : 5µL of NarG+1µL of 6X loading dye
- Well 7 : 5µL of NarG+1µL of 6X loading dye
- Gel : 1.2%
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Expected sizes :
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We obtain fragments at the right size. We can purify it.
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4 - Gel purification of PCR products : NarK, NarG, NirB
Xavier
Protocol : Gel purification
REPRIS dans 20µL !!!!!!!! ????????? PAS DE NANODROP ??????
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