|
|
Customer pHZ5 Based Plasmid
|
|
|
|
|
|
Transform RFY309
|
|
|
|
|
STOP
|
no  |
Transactivation Potential <~10-5
|
|
|
yes  |
|
|
Mate to Library Strain(s)
|
|
|
|
|
|
Select for Marker Expression
|
|
|
|
|
|
Screen for Glucose Repression and Second Marker Expression
|
|
|
|
|
Discard
|
no |
Correct Phenotype?
|
|
|
|
yes |
|
|
|
Colony PCR
|
|
|
|
|
|
|
|
Gap Repair (Recombination) Clone
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Go Back to Colony PCR
|
no |
Was Cloning Successful?
|
|
|
|
yes |
|
Discard
|
no |
Was Phenotype Recovered?
|
|
|
|
yes |
|
|
|
Sequence PCR Products
|
