Difference between revisions of "Team:Paris Saclay/Notebook/August/18"
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=Tuesday 18th August= | =Tuesday 18th August= | ||
==Lab Work== | ==Lab Work== | ||
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Incubation 37°C, 3h | Incubation 37°C, 3h | ||
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===Plasmid extraction=== | ===Plasmid extraction=== | ||
''by Pauline'' | ''by Pauline'' | ||
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* BBa_K1707022 #1 and #2 | * BBa_K1707022 #1 and #2 | ||
* BBa_K1707023 #1 and #2 | * BBa_K1707023 #1 and #2 | ||
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With Sylvie Lautru's Protocol | With Sylvie Lautru's Protocol | ||
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===Electrophoresis=== | ===Electrophoresis=== | ||
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Agarose gel 1%, migration 90V | Agarose gel 1%, migration 90V | ||
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* BBa_K1707035 #1 and #2 | * BBa_K1707035 #1 and #2 | ||
* BBa_K1707036 #1 and #2 | * BBa_K1707036 #1 and #2 | ||
| − | + | [[File:ParisSaclay 18.08.15-vérif.jpg|300px|center]] | |
| + | <html><p><i>Verification by digestion with XbaI and PstI, from left to right: 1. <a href="https://2015.igem.org/File:Paris_Saclay-Ladder.jpg" target="_blank">DNA Ladder</a>, 2. BBa_K1707035#1, 3. BBa_K1707035#2, 4. BBa_K1707036#1, 5. BBa_K1707036#2, 6. Empty, 7. Empty, 8. Empty, 9. Empty, 10. Empty</i></p></html> | ||
We can conclude that BBa_K1707035 #1, BBa_K1707036 #1 and #2 are OK, but not BBa_K1707035 #2 | We can conclude that BBa_K1707035 #1, BBa_K1707036 #1 and #2 are OK, but not BBa_K1707035 #2 | ||
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in 10mL LB without antibiotic | in 10mL LB without antibiotic | ||
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===PCR=== | ===PCR=== | ||
''by Audrey'' | ''by Audrey'' | ||
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* BBa_K1707000 #2, #3, #4 | * BBa_K1707000 #2, #3, #4 | ||
* BBa_K1707013 #1 | * BBa_K1707013 #1 | ||
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''by Pauline'' | ''by Pauline'' | ||
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* BBa_K1707022 #1 and #2 | * BBa_K1707022 #1 and #2 | ||
* BBa_K1707023 #1 and #2 | * BBa_K1707023 #1 and #2 | ||
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Incubation 37°C, 1h | Incubation 37°C, 1h | ||
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===Transplant=== | ===Transplant=== | ||
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===PCR=== | ===PCR=== | ||
''by Audrey'' | ''by Audrey'' | ||
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'''Member present:''' | '''Member present:''' | ||
Latest revision as of 21:10, 18 September 2015
Contents
Tuesday 18th August
Lab Work
Digestion
by Pauline
Biobricks:
- BBa_K1707035 #1 and #2
- BBa_K1707036 #1 and #2
Mix for each plasmid:
- 2 µL plasmid
- 0,5 µL XbaI
- 0,5 µL PstI
- 1 µL Buffer FastDigest 10x
- 6 µL H2O
Incubation 37°C, 3h
Plasmid extraction
by Pauline
- BBa_K1707022 #1 and #2
- BBa_K1707023 #1 and #2
- BBa_K1707034 #1 and #2
With Sylvie Lautru's Protocol
Electrophoresis
by Pauline
Agarose gel 1%, migration 90V
- BBa_K1707035 #1 and #2
- BBa_K1707036 #1 and #2
Verification by digestion with XbaI and PstI, from left to right: 1. DNA Ladder, 2. BBa_K1707035#1, 3. BBa_K1707035#2, 4. BBa_K1707036#1, 5. BBa_K1707036#2, 6. Empty, 7. Empty, 8. Empty, 9. Empty, 10. Empty
We can conclude that BBa_K1707035 #1, BBa_K1707036 #1 and #2 are OK, but not BBa_K1707035 #2Inoculation
by Pauline
3 strains: 1320; 1693; 1696
in 10mL LB without antibiotic
PCR
by Audrey
- BBa_K1707000 #2, #3, #4
- BBa_K1707013 #1
- BBa_K1707030 #1
- BBa_K1707019 #1
- BBa_K1707020 #2
- BBa_K1707035 #1
- BBa_K1707036 #1
- BBa_K1707027 #1
- BBa_K1707021 #2
Control +: BBa_K115017 Control -: BBA_J23101+GFP (Interlab study)
Vtot in each tube= 50µL: 2µL plasmid + 48µL mix
Mix for all tubs:
- 476,25 µL H2O
- 150 µL Buffer 5X
- 7,5 µL Forward Primer (iPS43)
- 7,5 µL Reverse Primer (iPS3)
- 15 µL dNTP
- 3,75 µL GoTAQ
- 60 µL MgCl2
Digestion
by Pauline
- BBa_K1707022 #1 and #2
- BBa_K1707023 #1 and #2
- BBa_K1707034 #1 and #2
Mix for each plasmid:
- 2 µL plasmid
- 0,5 µL XbaI
- 0,5 µL PstI
- 1 µL Buffer FastDigest 10x
- 6 µL H2O
Incubation 37°C, 1h
Transplant
by Pauline
Biobrick: BBa_K1707031 clone #7 to #26 In a liquid culture: 5mL LB + 5µL Antibiotic
PCR
by Audrey
Member present:
- Instructors: Claire
- Students: Pauline and Audrey
Contact
eigem.parissaclay AT gmail DOT com
t@iGEMParisSaclay
aUniversité Paris Sud
a91405 Orsay, France
Copyright © 2015 iGEM Paris Saclay. All rights reserved All inspirations encouraged.
*This wiki was conceived during long nights of work and a great coffee experience in the beautiful Orsay, France.
eigem.parissaclay AT gmail DOT com
t@iGEMParisSaclay
aUniversité Paris Sud
a91405 Orsay, France
Copyright © 2015 iGEM Paris Saclay. *This wiki was conceived during long nights of work and a great coffee experience in the beautiful Orsay, France.