Team:DTU-Denmark/Notebook/10 June 2013

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10 June 2013

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Contents

208 lab


Main purposes today


  • helloworld-project
  • OD meassurements
  • autoclaving
  • make competent cells

Who was in the lab


Kristian, Jakob, Henrike

Procedure


OD measurements


timeOD600 measurement
0.190
11:160.289
11:280.356
11:320.390
taken out 11:340.397

placed on ice

Making competent cells


Using the Protocol, we got from Andreas with following modifications to protocol:

  • made only half the amount
  • used falcon tubes

The competent cells were stored in -80 C freezer on the lower shelf

Autoclaving


following was autoclaved:

  • Eppendorf tubes
  • 3 x 100ml dm water
  • 2 x 100ml 0.1M CaCl_2 and 15% glycerol
  • 1 L bottle of 0.1M CaCl_2
  • 1 L bottle of 50% glycerol
  • 200ml bottle of 1M CaCl_2

Solutions made


  • 0.1 M CaCl_2 and 15% glycerol from 10ml 1M CaCl_2
  • 30ml 50% glycerol 70ml dm water
  • 0.1 M CaCl_2 from 100ml 1M CaCl_2 and 900ml dm water
  • chlorampenicol stock 1ml with cencentration 50mg/ml

Plates


  • 24 ps with 10ug/ml chlorampenicol (CAP) 5 mg to 500ml
  • 33 ps with 30ug/ml Kanamycin 0.3ml of 30ug/ml to 500ml

Transformation


Following the official iGEM protocol :

  • BBa-I0500 plate 5 well 14N, then moved to -80C
  • BBa-I14032 plate 3 well 18H, then moved to -80C

Transformation efficiency test (0.5,5,10,20,50)

Incubation 2 hours with 300 rpm

Plates:

  • I0500 - no incubation on Kanamycin plate, one 20ul and one 200 ul
  • I14032 - no incubation on CAP, one 20ul and one 200 ul

all 4 had 2 hours incubation.


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