Team:TU Darmstadt/Notebook/sec1/K1602033
K1602033 - B0034-Xylanase-PDZ-Ligand (B0034-xynA-PDZlig)
To connect LINK ZU REGISTRY B0034-XYNA to the in vitro scaffold a linker was added using a PCR with the oligonucleotides VF2 and XynA_fus_rev. The PCR product was verified by gel electrophoresis (Figure 1), digested with DpnI and purified. 1 ng of this product was used for a second PCR to add a ligand to the linker (Figure 2). The product was purified and cloned into pSB1C3, followed by a transformation of E. coli Top 10. Positive colonies were screened by CPCR (Figure 3).
Figure 1 Linker PCR of B0034-xynA (1). The size of the amplified product was around 0.8 kbp. DNA marker: Gene Ruler 1 kB DNA Ladder. (NEB).
Figure 2 PCR of B0034-xynA-PDZlig (1). The size of the amplified product was around 1.0 kbp. DNA marker: 2-Log DNA Ladder. (NEB).
Figure 3 Colony PCR of B0034-xynA-PDZlig (1). The size of the amplified product was around 1.0 kbp. DNA marker: Gene Ruler 1 kB DNA Ladder. (NEB).