Difference between revisions of "Team:TU Darmstadt/Notebook/sec1/K1602044"
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| − | <a href=" https://2015.igem.org/Team:TU_Darmstadt/Notebook/sec1/K1602041 " title="Opens internal link in current window" class="internal link">K1602041</a> was digested with <em>Xba</em>I and <em>Pst</em>I. The construct was cloned into the vector pSB1A2 containing T7 promoter and transformed into competent <em>E. coli</em> Top 10 via heat shock. Colonies were screened by colony PCR (oligonucleotides VF2 and VR) (Figure 1). Positive clones were inoculated and the plasmids were extracted. Afterwards the construct was cloned into the vector pSB1C3 using <em>EcoR</em>I and <em>Pst</em>I. The pSB1C3 vector containing the coding sequence for T7-B0034-YFP-SH3Lig was used for protein expression in <em>E. coli</em> strain BL21. | + | <a href=" https://2015.igem.org/Team:TU_Darmstadt/Notebook/sec1/K1602041 " title="Opens internal link in current window" class="internal link">K1602041</a> was digested with <em>Xba</em>I and <em>Pst</em>I. The construct was cloned into the vector pSB1A2 containing T7 promoter and transformed into competent <em>E. coli</em> Top 10 via heat shock. Colonies were screened by colony PCR (oligonucleotides VF2 and VR) (Figure 1). Positive clones were inoculated and the plasmids were extracted. Afterwards the construct was cloned into the vector pSB1C3 using <em>EcoR</em>I and <em>Pst</em>I. The pSB1C3 vector containing the coding sequence for T7-B0034-YFP-SH3Lig was used for protein expression in <em>E. coli</em> strain BL21. </p> |
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<img src=" https://static.igem.org/mediawiki/2015/f/f0/Bild_3%3B_Colony_PCR_of_T7-B0034-YFP-Linker-SH3.png" width=100% height=100%> | <img src=" https://static.igem.org/mediawiki/2015/f/f0/Bild_3%3B_Colony_PCR_of_T7-B0034-YFP-Linker-SH3.png" width=100% height=100%> | ||
<figcaption><br><b>Figure 1</b> Colony PCR of T7-B0034-YFP-SH3Lig (3-5). The size of the amplified product was around 1.1 kbp. DNA marker: 2-Log DNA Ladder. (NEB).</figcaption> | <figcaption><br><b>Figure 1</b> Colony PCR of T7-B0034-YFP-SH3Lig (3-5). The size of the amplified product was around 1.1 kbp. DNA marker: 2-Log DNA Ladder. (NEB).</figcaption> | ||
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Revision as of 18:07, 17 September 2015
K1602044 - T7-B0034-enhanced yellow fluorescent protein-SH3-Ligand (T7-B0034-eYFP-SH3Lig)
K1602041 was digested with XbaI and PstI. The construct was cloned into the vector pSB1A2 containing T7 promoter and transformed into competent E. coli Top 10 via heat shock. Colonies were screened by colony PCR (oligonucleotides VF2 and VR) (Figure 1). Positive clones were inoculated and the plasmids were extracted. Afterwards the construct was cloned into the vector pSB1C3 using EcoRI and PstI. The pSB1C3 vector containing the coding sequence for T7-B0034-YFP-SH3Lig was used for protein expression in E. coli strain BL21.
Figure 1 Colony PCR of T7-B0034-YFP-SH3Lig (3-5). The size of the amplified product was around 1.1 kbp. DNA marker: 2-Log DNA Ladder. (NEB).