Team:Paris Saclay/Notebook/July/21

From 2014.igem.org

(Difference between revisions)
(1 - Results: Transformation of supercompetent cells with CaCl2)
(Lab Work)
Line 12: Line 12:
Results: Nothing has grown.
Results: Nothing has grown.
-
===2 - Oligo's design===
+
===2 - Results: Transformation of DY330 via pJBEI6409===
 +
 
 +
''by Sean''
 +
 
 +
Transformation performed on the [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/18 18th July]
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{| class="wikitable"
 +
|+Number of colonies per dish
 +
|-
 +
! 50μL from cuvette
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! 100μL
 +
! remainder
 +
|-
 +
| 0
 +
| 4
 +
| 30
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! 2μL of plasmid
 +
|-
 +
| 3
 +
| 6
 +
| 45
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! 4μL
 +
|-
 +
| 0
 +
| 0
 +
| 0
 +
!Control
 +
|}
 +
 
 +
 
 +
===3 - Oligo's design===
''by Romain''
''by Romain''
-
===3 - Liquid Bacterial Culture===
+
===4 - Liquid Bacterial Culture===
''by Marie, Romain & Sean''
''by Marie, Romain & Sean''
Line 24: Line 54:
*The new strains received
*The new strains received
-
===4 - Electrophoresis===
+
===5 - Electrophoresis===
''by Fabio (process A) and Mathieu (process B and C)''
''by Fabio (process A) and Mathieu (process B and C)''

Revision as of 21:03, 21 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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