Team:Paris Saclay/Notebook/July/21

From 2014.igem.org

(Difference between revisions)
(2 - Results: Transformation of DY330 via pJBEI6409)
(Monday 21st July)
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===1 - Results: Transformation of supercompetent cells with CaCl<sub>2</sub>===
===1 - Results: Transformation of supercompetent cells with CaCl<sub>2</sub>===
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''by Romain''
''by Romain''
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===2 - Results: Transformation of DY330 via pJBEI6409===
===2 - Results: Transformation of DY330 via pJBEI6409===
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''by Sean''
''by Sean''
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|+Number of colonies per dish
|+Number of colonies per dish
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! 50μL from cuvette
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! 50μl from cuvette
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! 100μL
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! 100μl
! remainder  
! remainder  
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| 30
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! 2μL of plasmid
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! 2μl of plasmid
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! 4μL
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! 4μl
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===3 - Oligo's design===
===3 - Oligo's design===
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''by Romain''
''by Romain''
===4 - Liquid Bacterial Culture===
===4 - Liquid Bacterial Culture===
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''by Marie, Romain & Sean''
''by Marie, Romain & Sean''
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===5 - Electrophoresis===
===5 - Electrophoresis===
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''by Fabio (process A) and Mathieu (process B and C)''
''by Fabio (process A) and Mathieu (process B and C)''
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[[File:LU000070.jpg|right]]
[[File:LU000070.jpg|right]]
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We used 2µL of DNA of the following Biobricks in a 1% Agarose Gel.
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We used 2µl of DNA of the following Biobricks in a 1% Agarose Gel.
''The PCRs came from Mathias' manipulation made the [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/18 18th July].''
''The PCRs came from Mathias' manipulation made the [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/18 18th July].''

Revision as of 13:49, 22 July 2014

Contents

Monday 21st July

Lab Work

1 - Results: Transformation of supercompetent cells with CaCl2

by Romain

Strains transformed: MG1655, MG1655Z1. from Bacterial Cultures transformed the 18th July

Results: Nothing has grown.

2 - Results: Transformation of DY330 via pJBEI6409

by Sean

Transformation performed on the 18th July

Number of colonies per dish
50μl from cuvette 100μl remainder
0 4 30 2μl of plasmid
3 6 45 4μl
0 0 0 Control

3 - Oligo's design

by Romain

4 - Liquid Bacterial Culture

by Marie, Romain & Sean

  • DY330 pJBEI6409 with 10µl Cm in 10ml LB (x2)
  • BT340 Cm and Amp
  • The new strains received

5 - Electrophoresis

by Fabio (process A) and Mathieu (process B and C)

LU000069.jpg
LU000071.jpg
LU000070.jpg

We used 2µl of DNA of the following Biobricks in a 1% Agarose Gel. The PCRs came from Mathias' manipulation made the 18th July.

Process A

  1. J23119 Cl1
  2. J23119 Cl2
  3. J23106 Cl1
  4. J23100 Cl2
  5. PCR 1
  6. PCR 2
  7. PCR 3
  8. PCR 4
  9. PCR 5
  10. PCR 6
  11. PCR 7
  12. PCR 8
  13. PCR 9
  14. PCR 10

Process B

  1. J23100 Cl1
  2. J23100 Cl2
  3. J23106 Cl1
  4. J23106 Cl2
  5. J23114 Cl1
  6. J23114 Cl2

Process C

Pooling and purifying PCR 9 and 10 from process A.

  1. PCR purified products. IPS70 / IPS71 of pOsV230 (Apramycin)
  2. BT 340 (plasmid's flipase)

Results:

  • A: From 1 to 4: Success, DNAs have the expected size.
  • A: From 5 to 12: Failure, No PCR products.
  • A: Numbers 13 and 14: Success, PCR products have the expected size.
  • B: All 6 extractions were successful.
  • C: Number 1: successful concentration of pOsV230's PCR product.
  • C: Number 2 had no migration.

Protocol

People there:

  • Instructors and advisors: Solenne and Sylvie.
  • Students: Arnaud, Fabio, Juliette, Mathieu, Marie, Pierre, Romain, Sean and Terry.

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