Vector Digestions
From 2006.igem.org
(Difference between revisions)
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# 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 | + | (Todd -- I think the guys -- Brad, Eric, Kelly, Adam -- finished this this morning, but they did not document it. I was in class and they are gone now. My understanding is that they plan to have everyone here at noon tomorrow. They will hopefully be IM able to communicate with you then if needed.) |
Revision as of 19:00, 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 and they are gone now. My understanding is that they plan to have everyone here at noon tomorrow. They will hopefully be IM able to communicate with you then if needed.)
