Figure 1 PCR of cadA (X). The size of the amplified product was around 1.5 kbp. DNA marker: 2-Log DNA Ladder (NEB).
The xylC-gene was synthesized by GeneArt and amplified
through PCR using the xylC (FW) and xylC (REV)
oligonucleotides. The PCR product was verified by agarose gel
electrophoresis, digested with DpnI for 120 minutes and subsequently
purified. The xylC amplicon was cut with EcoRI and PstI and
ligated into a pSB1C3 vector using T4-ligase. E.coli Top 10
has been transformed with the ligation product by heat shock
transformation and the cells were spread out on a CMP agar plate.
Clones were screened by colony PCR using VF2 and VR oligonucleotides.
Plasmid DNA was isolated from positive clones which were verified by
sanger sequencing.
