Team:TU Darmstadt/Notebook/sec1/K1602056
K1602056 - B0034-D-Xylonic Acid Dehydratase and B0034-Aldehyde Reductase (B0034-yjhG-B0034-yqhD)
Figure 1 Restriction digest of B0034-xylB-B0034-xylC (along with others). You can see the xylB-xylC string (below 2 kbp) and the pSB1C3 backbone (above 2 kbp). DNA marker: 2-Log DNA Ladder (NEB)..
The Bba_K1602002 part and the Bba_K1602015 part were combined.
Firstly the B0034-yjhG composite was digested with EcoRI and SpeI while the B0034-YqhD composite was digested with XbaI and PstI. They both were taken from a pSB1A2 plasmid. The restriction products were dephosphorylated and ligated in one step into a pSB1C3 vector, which was cut with EcoRI and PstI. E. coli Top 10 cells were transformed with the ligation product. The resulting colonies were screened for positive clones by restriction digest with EcoRI and PstI. Plasmid DNA was isolated from positive clones and verified by sanger sequencing.