Team:TU Darmstadt/Notebook/sec1/K1602010

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K1602010 - D-Xylonolactonase (xylC)

Figure 1 PCR of xylC. The size of the amplified product was 1 kbp. DNA marker: 2-Log DNA Ladder (NEB).

The xylC-gene was synthesized by GeneArt and amplified by 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.