IGEM:UC Berkeley/2006/Conjugation
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< IGEM:UC Berkeley/2006(Difference between revisions)
Current revision (14:33, 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> | ||
| - | [[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 | ||
Current revision
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
