Difference between revisions of "Team:TU Darmstadt/Notebook/sec1/K1602056"
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| − | <img class="transparent" alt="https://static.igem.org/mediawiki/2015/ | + | <img class="transparent" alt="https://static.igem.org/mediawiki/2015/0/02/TU_Darmstadt_Gel_B0034-yjhG-B0034-yqhD.png" src="https://static.igem.org/mediawiki/2015/0/02/TU_Darmstadt_Gel_B0034-yjhG-B0034-yqhD.png" height="250" width="150"> |
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| + | <p align="justify"><b>Figure 1</b> Gel of CPCR. B0034-<i>yjhG</i>-B0034-<i>yqhD</i> string has a length of approximately 3 kbp if you use VF2 and VR oligonucleotides. DNA marker: 2-Log DNA Ladder (NEB).</p>.</figcaption> | ||
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| − | The | + | The <a href=" https://2015.igem.org/Team:TU_Darmstadt/Notebook/sec1/K1602002 " title="Opens internal link in current window" class="internal link">BBa_K1602002</a> part and the <a href=" https://2015.igem.org/Team:TU_Darmstadt/Notebook/sec1/K1602015 " title="Opens internal link in current window" class="internal link">BBa_K1602015</a> part were combined.</p> |
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| − | Firstly the B0034-<em>yjhG</em> composite was digested with <em>EcoR</em>I and <em>Spe</em>I while the B0034-<em>YqhD</em> composite was digested with <em>Xba</em>I and <em>Pst</em>I. They both were taken from a pSB1A2 plasmid. The restriction products were dephosphorylated and ligated in one step into a pSB1C3 vector, which was cut with <em>EcoR</em>I and <em>Pst</em>I. <em>E. coli</em> Top 10 cells were transformed with the ligation product. The resulting colonies were screened for positive clones by | + | Firstly the B0034-<em>yjhG</em> composite was digested with <em>EcoR</em>I and <em>Spe</em>I while the B0034-<em>YqhD</em> composite was digested with <em>Xba</em>I and <em>Pst</em>I. They both were taken from a pSB1A2 plasmid. The restriction products were dephosphorylated and ligated in one step into a pSB1C3 vector, which was cut with <em>EcoR</em>I and <em>Pst</em>I. <em>E. coli</em> Top 10 cells were transformed with the ligation product. The resulting colonies were screened for positive clones by colony PCR using VR and VF2 oligonucleotides. Plasmid DNA was isolated from positive clones and verified by sanger sequencing.</p> |
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Latest revision as of 20:11, 18 September 2015
K1602056 - B0034-D-Xylonic Acid Dehydratase and B0034-Aldehyde Reductase (B0034-yjhG-B0034-yqhD)
Figure 1 Gel of CPCR. B0034-yjhG-B0034-yqhD string has a length of approximately 3 kbp if you use VF2 and VR oligonucleotides. DNA marker: 2-Log DNA Ladder (NEB).
.The BBa_K1602002 part and the BBa_K1602015 part were combined.
Firstly the B0034-yjhG composite was digested with EcoRI and SpeI while the B0034-YqhD composite was digested with XbaI and PstI. They both were taken from a pSB1A2 plasmid. The restriction products were dephosphorylated and ligated in one step into a pSB1C3 vector, which was cut with EcoRI and PstI. E. coli Top 10 cells were transformed with the ligation product. The resulting colonies were screened for positive clones by colony PCR using VR and VF2 oligonucleotides. Plasmid DNA was isolated from positive clones and verified by sanger sequencing.