Vector Digestions
From 2006.igem.org
Prepare linearized vector from pTet-GFP in the following two ways:
- cut with XbaI + SpeI in order to receive PCR product of reversed pBAD
- cut with EcoRI + PstI in order to receive annealed hix oligos
Each of the above should be done this way:
- put together a restriction digestion using 400 ng vector and 40 ul volume
- run on 1% agarose gel
- purify the 2200 bp fragment (there will also be a smaller fragment from the pTet-GFP)