Team:DTU-Denmark/Notebook/10 June 2013

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==Procedure==
==Procedure==
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===OD measurements===
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{|
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!time!!OD600 measurement
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|-
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| ||0.190
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|-
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|11:16||0.289
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|-
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|11:28||0.356
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|-
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|11:32||0.390
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|-
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|taken out 11:34||0.397
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|}
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placed on ice
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===Making competent cells===
===Making competent cells===
Using the [[Team:DTU-Denmark/Methods/Competent Cells|Protocol]], we got from Andreas with following modifications to protocol:
Using the [[Team:DTU-Denmark/Methods/Competent Cells|Protocol]], we got from Andreas with following modifications to protocol:

Revision as of 15:47, 25 June 2013

Contents

main purposes today

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

who were 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 [http://parts.igem.org/Help:Protocols/Transformation 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.



Conclusion from today

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