Difference between revisions of "Team:TU Darmstadt/Notebook/sec1/K1602024"

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<h3>K1602024 - HistidinTag-Green fluorescent protein-SilicaTag4 (His-GFP-Si4)</h3>
  
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<a href=” https://2015.igem.org/Team:TU_Darmstadt/Notebook/sec1/K1602021”title="Opens internal link in current window" class="internal link">K1602021<span        style="white-space: pre;"> -&nbsp;</span><span style="white-space: pre;">HistidinTag-Green fluorescent protein (</span>His-GFP)</a> 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 <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. </p>
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<figcaption><br><b>Figure 1</b> SOE PCR of His-GFP (1). The size of the amplified product 1 was around 0.9 kbp. DNA marker: 2-Log DNA Ladder.
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<figcaption><br><b>Figure 2</b> 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.
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Revision as of 20:26, 17 September 2015

K1602024 - HistidinTag-Green fluorescent protein-SilicaTag4 (His-GFP-Si4)

K1602021 - HistidinTag-Green fluorescent protein (His-GFP) 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.