Difference between revisions of "Team:Pasteur Paris/Week 1"

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Revision as of 12:28, 18 August 2015

WEEK 1

06/01 - 06/05

WEEK 2

06/08 - 06/12

WEEK 3

06/15 - 06/19

WEEK 4

06/22 - 06/26

WEEK 5

06/29 - 07/03

WEEK 6

07/06 - 07/10

WEEK 7

07/13 - 07/17

WEEK 8

07/20 - 07/24

WEEK 9

07/27 - 07/31

WEEK 10

08/03 - 08/07

WEEK 11

08/10 - 08/14

WEEK 12

08/17 - 08/21

WEEK 13

08/24 - 08/28

WEEK 14

08/31 - 09/04

WEEK 15

09/07 - 09/11

WEEK 1

06/01 - 06/05

04-06-15 : Transformation of our biobricks in DH5-alpha competent cells : xxxxxxxx

    Materials :
  • Competant DH5-alpha cells (100 microL per transformation)
  • Ice (in ice bucket/container)
  • 2mL tube (1 per a transformation)
  • 42°C water bath
  • SOC media
  • Petri dishes xith LB agar and appropriate antibiotic (2 per transformation)
  • 37°C incubator

    Protocol :
  1. Thaw on ice one tube of DH5α cells. Place 1.5mL microcentrifuge tubes on wet ice.
  2. Gently mix cells with the pipette tip and aliquot 50µL for each transformation into a 1.5ml centrifuge tube.
  3. Refreeze any unused cells.
  4. Add 1 to 5 µL (1-10 ng) DNA to the cell and mix gently (Not mix by pupetting up and down)
  5. Incubate tubes on ice for 30 min
  6. Heat shock cells for 40s in a 42°C water bath without shaking.
  7. Place tube on ice for 3 minutes.
  8. Add pre-warmed 700µL SOC medium (XXXXXXXX).
  9. Incubate at 37°C for 40 min under shaking condition (200 rpm)
  10. Spread 200 µL of transformed cell on LB + Cm34 (chloramphenicol 34µg/mL).
  11. Incubate dished overnight at 37°C


05-06-15 : Bacterial amplification : xxxxxxxxxxx Materials : • Petri dishes • LB with or without chlroramphénicol • 15mL falcon tubes (3 per Petridish used) Protocol : 1/ Fill the tubes with 5mL of the correct media (3 tubes per strain) 2/ Pick three transformed clones per Petri box (1 clone per 15mL tube) 3/ Incubate in agitation (8 hours at 200 rpm, 37°C)

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