IGEM:UC Berkeley/2006/Conjugation

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Current revision (13:33, 17 July 2006) (view source)
 
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[[IGEM:UC Berkeley/2006/Rlambda|Rlambda (marker swap Procedure)]]<br>
[[IGEM:UC Berkeley/2006/Rlambda|Rlambda (marker swap Procedure)]]<br>
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[[IGEM:UC Berkeley/2006/TlambdaOlambda|TlambdaOlambda]]
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[[IGEM:UC Berkeley/2006/TlambdaOlambda|TlambdaOlambda]] <br>
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== protocol for conjugation ==
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'''donor protocol'''
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#start out with saturated 1 mL of culture
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#spin down (top speed for 10 sec.) , discard fluid
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# resuspend in 1 mL LB
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#vortex, spin, discard fluid
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#resuspend in 1 mL LB, vortex
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#put 200uL of culture into 2 mL of LB  (if arabinose is needed put in 100 mg/mL)
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# grow 2 hours
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# do Assay
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'''Recipient protocol'''
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#take aliquot and thaw
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#dilute 10 fold into LB (ie. put 200 uL into 2 mL of LB)
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#grow 2 hours
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#do Assay

Current revision

Rlambda (marker swap Procedure)
TlambdaOlambda

protocol for conjugation

donor protocol

  1. start out with saturated 1 mL of culture
  2. spin down (top speed for 10 sec.) , discard fluid
  3. resuspend in 1 mL LB
  4. vortex, spin, discard fluid
  5. resuspend in 1 mL LB, vortex
  6. put 200uL of culture into 2 mL of LB (if arabinose is needed put in 100 mg/mL)
  7. grow 2 hours
  8. do Assay

Recipient protocol

  1. take aliquot and thaw
  2. dilute 10 fold into LB (ie. put 200 uL into 2 mL of LB)
  3. grow 2 hours
  4. do Assay
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