1255 pTXB1-C6H2
Cloning of 1255 pTXB1-C6H2
[edit]Purpose: Cloning of 1255 pTXB1-C6H2
Date: July 6, 2012
Notebook:
Page:
Clone ID: 1255
Experimenters: Jennifer Truong
Results
[edit]Cloning
[edit]Amplified from 1126 (pTXB1) template with pTXB1-Xho1-F and pTXB1-Pst-R2 primers
Cut out band in lane 4, expected size 624 bp, purified and digested with restriction enzymes Xho1 and Pst1
Digested DNA and vector with restriction enzymes Xho1 and Pst1
Cut out bands from lane 3 and 4, purify, ligate and transform, plate and let grow overnight
Is lane 4 vector pTXB1? it should have two bands, one vector, one original insert.
Dropped DNA band, and vector band is a little too light, reamplified
Cut out lane 1 (using Phusion from Thermo) and 2 (using homemade Phusion), expected size 624 bp, purified and digested with restriction enzymes Xho1 and Pst1,
Lane 1 and 2 are the digested DNA and vector with Xho1+ Pst1 (from Phusion from Thermo)
Lane 3 and 4 are the digested DNA and vector with Xho1 and Pst1 (from Homemade Phusion)
Lane 4 digestion does not look good (vector)
Only cut out lane 1 and 2, purified and ligated together overnight. Paul transformed with Top10 cells and plated
Colony Screening
[edit]Date: June 9, 2012
Digested MiniPrep plasmids with restriction enzymes Xho1 and Pst1
May have postive insert for all samples, but unclear