Vector Digestions
From 2006.igem.org
(Difference between revisions)
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# run on 1% agarose gel | # run on 1% agarose gel | ||
# purify the 2200 bp fragment (there will also be a smaller fragment from the pTet-GFP) | # purify the 2200 bp fragment (there will also be a smaller fragment from the pTet-GFP) | ||
| + | |||
| + | (Todd -- I think the guys -- Brad, Eric, Kelly, Adam -- finished this this morning, but they did not document it. I was in class by the time they left and they are gone. My understanding is that they plan to have everyone here at noon tomorrow.) | ||
Revision as of 18:59, 19 June 2006
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)
(Todd -- I think the guys -- Brad, Eric, Kelly, Adam -- finished this this morning, but they did not document it. I was in class by the time they left and they are gone. My understanding is that they plan to have everyone here at noon tomorrow.)
