Team:TU Darmstadt/Notebook/sec1/K1602031
K1602031 – B0034-HistidinTag-Scaffold-SilicaTag4 (B0034-His-Scaffold-Si4)
K1602028 was used as template for PCR with oligonucleotides VF2 and Sc-Si4-REV1. After digesting the PCR product using DpnI and purification the silica-tag Si4 was connected to the scaffold by another PCR using the oligonucleotides VF2 and Sc-Si4-REV2. A gel extraction was performed. The purified PCR product was then digested with EcoRI and PstI. The construct was ligated into pSB1C3 and transformed into E. coli Top 10 via heat shock. Colonies were screened by colony PCR with VF2 and VR.