>You need SalI cut sites in the plasmid used for religation.
Clearly.
>Without GE, how do you know which segment of the cut dna you are
using? You need to purify the DNA section containing the light gene.
Could it work without purification? Maybe, but it might be harder to
do.
As I see it, there are 4 [EDIT: no, 6] possible outputs:
Ampicillin self ligated (will be destroyed by kanamycin)
Amp Lux (original) (destroyed)
Lux - ligated (destroyed, no ORI)
Kanamycin + Amp Ressistant
Kanam - Self ligated
Kanam + pVIB
Three of them will survive in selection medium. 1 out of three will glow. This one I'll take and streak out on another pate, ( and do a miniprep later)
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