Team:TU Darmstadt/Notebook/sec1/K1602024
K1602024 - HistidinTag-Green fluorescent protein-SilicaTag4 (His-GFP-Si4)
K1602021 was used as template for SOE-PCR with oligonucleotides VF2 and 33 (Figure 1). The silica-tag Si4 was added using another SOE-PCR with oligonucleotides VF2 and VR (Figure 2). The PCR product was purified and 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.
Figure 1 SOE PCR of His-GFP (1). The size of the amplified product 1 was around 0.9 kbp. DNA marker: 2-Log DNA Ladder.
Figure 2 SOE PCR of His-GFP-Si4 (1+2). The size of the amplified product 2 was around 1.1 kbp. DNA marker: 2-Log DNA Ladder.