Team:Paris/July 13

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Contents

Preparation of Competent Cells

  • Dilution 1/200e (1mL of the O/N culture on 200mL of LB)
  • Culture at 37°C until DO=0,57 (around 3h)
  • Sharing into 4 Falcon tubes of 50mL and cooling at 4°C during 40' in ice
  • Spin during 8', 2000rpm, 4°C
  • Resuspension of the cell's pellet in 20mL of cold CaCl2 (0,1M) during 30' in ice
  • Spin during 8', 2000rpm, 4°C
  • Resuspension of the cell's pellet in 1mL of cold CaCl2 (0,1M)
  • 1h of delay unexpected (problem with DNA lyophilized)
  • Transformation of the Biobricks with the "classical" protocol (--> heat shock at 42°C during 40")
  • Incubation O/N at 37°C of the transformed bacteria


Remark

  • Biobricks 2007 : use of 3µl of the minipreps
  • Biobricks 2008 : use of the maximum recoverable volume of the minipreps


List of the transformations

  • Biobricks 2007
Miniprep Details Results
MP1 J23100 Transformation
MP2 J23107 Transformation
MP3 B0015 Transformation
MP5 JB0030 Transformation
MP6 E0422 Transformation
  • Biobricks 2008
Ref. Book Plasmid Antibio Plate 2007 Results
2E_SP 1000 B0034 A 3O P1 No transformation
10F_SP 1001 R0079 A 19N P1 No transformation
4F_SP 1013 C0079 K 7K P2 No transformation
6H_SP 1001 C0179 A 6H P1 No transformation
6D_SP 1007 S03879 A / No transformation
6B_SP 1001 S03154 A 24K P1 No transformation
10F_SP 1002 J23101 A 21G P3 No transformation
11F_SP 1002 J23109 A 23G P3 No transformation
4C_SP 1000 R0040 A 7O P1 No transformation
2G_SP 1000 C0040 A 5C P1 No transformation
1C_SP 1008 PSB1A3 A 23E P1 No transformation
7G_SP 1013 E1010 K 15M P2 No transformation
7E_SP 1001 E0040 A 5H P1 No transformation
2F_SP 1016 E0030 K 7G P1 No transformation


Stock solution

  • TE (10:1) pH 8.0
  • plates of Ampicilin and Kanamycin
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