Bba_K1602025 was used as template for SOE-PCR with oligonucleotides VF2 and 33. The silica-tag Si4 was added using another SOE-PCR with oligonucleotides VF2 and VR. A PCR with oligonucleotides 1 and VR added the RBS to the construct. The PCR product was purified and digested with <em>EcoR</em>I and <em>Pst</em>I. The construct was ligated into pSB1C3 and transformed into <em>E. coli</em> Top 10 via heat shock. Colonies were screened by colony PCR with VF2 and VR. The plasmids were extracted and digested with <em>Xba</em>I and <em>Pst</em>I and ligated into B0034-pSB1A2 and transformed into <em>E. coli</em> Top 10. Colonies were screened by colony PCR with oligonucleotides VF2 and VR. Positive colonies were inoculated and the plasmids were extracted
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Bba_K1602027 was digested with <em>Xba</em>I and <em>Pst</em>I and ligated into B0034-pSB1A2 and transformed into <em>E. coli</em> Top 10. Colonies were screened by colony PCR with oligonucleotides VF2 and VR. Positive colonies were inoculated and the plasmids were extracted
<figcaption><br><b>Figure 1</b> SOE PCR of Prefix-His-Tag-GFP (1). The size of the amplified product 1 was around 0.9 kbp. DNA marker: Quick-Load Purple 2-Log DNA Ladder.
<figcaption><br><b>Figure 2</b> SOE PCR of Prefix-His-Tag-GFP-Si4 (1+2). The size of the amplified product 2 was around 1.1 kbp. DNA marker: Quick-Load Purple 2-Log DNA Ladder.
Bba_K1602027 was digested with XbaI and PstI and ligated into B0034-pSB1A2 and transformed into E. coli Top 10. Colonies were screened by colony PCR with oligonucleotides VF2 and VR. Positive colonies were inoculated and the plasmids were extracted
Figure 3 Colony PCR of B0034-Prefix-His-Tag-GFP-Si4 (1). Colony 1 contained the insert of around 1.1 kbp. DNA marker: Quick-Load Purple 2-Log DNA Ladder.
