Team:Paris Saclay/Notebook/July/24

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(Difference between revisions)
(→Results of the PCR Targeting)
(→Transformation of competent E.coli cells)
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# BBa_K228001 Undiluted
# BBa_K228001 Undiluted
# DH5α Control
# DH5α Control
 +
 +
'''Results on 25th July morning:'''
 +
 +
# Success, grown very well (> 200 colonies)
 +
# Success, grown well (> 50 colonies)
 +
# Success, grown well (> 90 colonies)
 +
# Success, grown (15 colonies)
 +
# Success, nothing has grown
[https://2014.igem.org/Team:Paris_Saclay/Protocols/Transformation_of_supercompetent_Ecoli_cells_with_CaCl2 Protocol]
[https://2014.igem.org/Team:Paris_Saclay/Protocols/Transformation_of_supercompetent_Ecoli_cells_with_CaCl2 Protocol]

Revision as of 14:55, 25 July 2014

Contents

Thursday 24th July

Lab work

B - Construction of the fusion protein (color)

Plasmid DNA Extraction and Electrophoresis

by Terry

Electrophoresis 24.07.14.jpg

Biobricks used:

  • 1. BBa_K1033927 with pSB1C3 (pink chromoprotein)
  • 2. BBa_K1033910 with pSB1C3 (yellow chromoprotein)
  • 3. BBa_K1033921 with pSB1C3 (red chromoprotein)
  • 4. BBa_K1033922 with pSB1C3 (red chromoprotein)
  • 5. BBa_K1033902 with pSB1C3 (blue chromoprotein)
  • 6. BBa_K731201 with pSB1C3 (Arabinose inductible araC-pBAD promoter)
  • 8. BBa_K1033925 with pSB1C3 (pink chromoprotein)
  • 9. BBa_K1033905 with pSB1C3 (purple chromoprotein)
  • 10. BBa_K1033913 with pSB1C3 (orange chromoprotein)
  • 11. BBa_K1033929 with pSB1C3 (blue chromoprotein)

Protocol

C - Lemon scent

Plasmid DNA Extraction & Electrophoresis

by Terry

Biobricks used:

  • 7. BBa_K762100 with pSB1C3 (Limonen synthase1 with RBS)

Protocol

Gel electrophoresis of p cola

LU000081.jpg

by Sean

p cola used: sample left overnight yesterday resuspended in 30μl of H2O milliQ.

Protocol

Results

Results of the PCR Targeting

by Romain

PCR targeting made the July 22nd. Strains used: DY330 (clone I and II). Plasmid used: pJBEI-6409 (code for enzymes to produce limonene from glucose in E. coli)

Results:

  • Nothing has grown on the control dishes.
  • Failure, nothing has grown on the clone I dishes.
  • Failure, other bacteria have grown on the clone II dishes.

We have to make another PCR. Liquid culture of DY330 made in 30ml LB.

E - Salicylate Inducible Suppressing System

Plasmid DNA Extraction & Electrophoresis

by Terry

Biobricks used:

  • 12 & 13. BBa_J45017 with pSB1C3 with pSB1AK3 (pchBA synthase (converts chorismate to salicylate))

Protocol

Transformation of competent E.coli cells

by Fabio

We transformed in DH5α both BioBricks rehydrated yesterday (BBa_J61051 and BBa_K228001).

Samples:

  1. BBa_J61051 Concentrated
  2. BBa_J61051 Undiluted
  3. BBa_K228001 Concentrated
  4. BBa_K228001 Undiluted
  5. DH5α Control

Results on 25th July morning:

  1. Success, grown very well (> 200 colonies)
  2. Success, grown well (> 50 colonies)
  3. Success, grown well (> 90 colonies)
  4. Success, grown (15 colonies)
  5. Success, nothing has grown

Protocol

Members present:

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

Back to the calendar

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