IGEM:UC Berkeley/2006/Conjugation: Difference between revisions
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[[IGEM:UC Berkeley/2006/Rlambda|Rlambda (marker swap Procedure)]]<br> | [[IGEM:UC Berkeley/2006/Rlambda|Rlambda (marker swap Procedure)]]<br> | ||
[[IGEM:UC Berkeley/2006/TlambdaOlambda|TlambdaOlambda]] | [[IGEM:UC Berkeley/2006/TlambdaOlambda|TlambdaOlambda]] <br> | ||
== protocol for conjugation == | |||
'''donor protocol''' | |||
#start out with saturated 1 mL of culture | |||
#spin down (top speed for 10 sec.) , discard fluid | |||
# resuspend in 1 mL LB | |||
#vortex, spin, discard fluid | |||
#resuspend in 1 mL LB, vortex | |||
#put 200uL of culture into 2 mL of LB (if arabinose is needed put in 100 mg/mL) | |||
# grow 2 hours | |||
# do Assay | |||
'''Recipient protocol''' | |||
#take aliquot and thaw | |||
#dilute 10 fold into LB (ie. put 200 uL into 2 mL of LB) | |||
#grow 2 hours | |||
#do Assay | |||
Latest revision as of 11:33, 17 July 2006
Rlambda (marker swap Procedure)
TlambdaOlambda
protocol for conjugation
donor protocol
- start out with saturated 1 mL of culture
- spin down (top speed for 10 sec.) , discard fluid
- resuspend in 1 mL LB
- vortex, spin, discard fluid
- resuspend in 1 mL LB, vortex
- put 200uL of culture into 2 mL of LB (if arabinose is needed put in 100 mg/mL)
- grow 2 hours
- do Assay
Recipient protocol
- take aliquot and thaw
- dilute 10 fold into LB (ie. put 200 uL into 2 mL of LB)
- grow 2 hours
- do Assay
