Team:Paris Saclay/Notebook/July/21

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'''Conclusion: The increase in colony number is proportional to the increase in volume used. The control yielded nothing. The results are coherent.'''
'''Conclusion: The increase in colony number is proportional to the increase in volume used. The control yielded nothing. The results are coherent.'''
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===3 - Oligonucleotide's design===
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==<span style="color:blue">D - Banana scent</span>==
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===1 - Oligonucleotide's design===
''by Romain''
''by Romain''

Revision as of 20:24, 23 July 2014

Contents

Monday 21st July

Lab Work

1 - Results: Transformation of CaCl2 supercompetent cells

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 (source of bacteria: electroporation cuvettes prepared on the 18th July)
Volume of plasmid used 50μl from cuvette 100μl remainder (850µl)
2μl of plasmid 0 4 30
4μl 3 6 45
Control 0 0 0

Conclusion: The increase in colony number is proportional to the increase in volume used. The control yielded nothing. The results are coherent.

D - Banana scent

1 - Oligonucleotide's design

by Romain

For the banana scent, design of iPS91 and iPS92, the ilvE sequence to transformed the Leucin into α-cetoisocaproate (The first step).

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. BBa_J23119 Cl1
  2. BBa_J23119 Cl2
  3. BBa_J23106 Cl1
  4. BBa_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. BBa_J23100 Cl1
  2. BBa_J23100 Cl2
  3. BBa_J23106 Cl1
  4. BBa_J23106 Cl2
  5. BBa_J23114 Cl1
  6. BBa_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.

Back to the calendar

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