Berk2010-Tim: Difference between revisions
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==[[User:Tim Hsiau|Tim Hsiau]] 15:19, 7 June 2010 (EDT)== | |||
Need to make MG1655 comp cells | |||
==[[User:Tim Hsiau|Tim Hsiau]] 20:13, 4 June 2010 (EDT)== | ==[[User:Tim Hsiau|Tim Hsiau]] 20:13, 4 June 2010 (EDT)== | ||
Talked to Jin about payload delivery device | Talked to Jin about payload delivery device |
Revision as of 12:19, 7 June 2010
Tim Hsiau 15:19, 7 June 2010 (EDT)
Need to make MG1655 comp cells
Tim Hsiau 20:13, 4 June 2010 (EDT)
Talked to Jin about payload delivery device
first column is PDD: payload delivery device
second column is the payload plus some stuff that may inhibit vacuole merging (secretion system)
third column is the BAC that contains other secretion system components
PDD's are not stable, need to make competent cells of BAC plus payload and do fresh transforms of PDD's everytime you assay
We have to work with MG1655 for assays, other strains okay for cloning
Other notes:
Mg needs to be added for all steps of cloning the PDD's 1M MgSO4 10µL into rescue step (after heatshock) 400µL spread on plate (if +Spec, add 20µL and spread) 30µL into ea mL TB media (use TB for growing up for assay)