Difference between revisions of "Team:TU Darmstadt/Notebook/sec1/K1602056"
(Created page with "{{:Team:TU_Darmstadt/Templates/CSS}} <h3>K1602016 - B0034-<small>D</small>-Xylose Dehydratase Plus B0034-<small>D</small>-Xylonolacton Lactonase (B0034-xylB-B0034-xylC)</h3>...") |
|||
| Line 1: | Line 1: | ||
{{:Team:TU_Darmstadt/Templates/CSS}} | {{:Team:TU_Darmstadt/Templates/CSS}} | ||
| − | <h3> | + | <h3>K1602056 - B0034-<small>D</small>-Xylonic Acid Dehydratase and B0034-Aldehyde Reductase (B0034-yjhG-B0034-yqhD)</h3> |
<html><div class="contentSection"> | <html><div class="contentSection"> | ||
<figure class="rightFig" style="width:55%"> | <figure class="rightFig" style="width:55%"> | ||
| − | <img class="transparent" alt="https://static.igem.org/mediawiki/2015/e/eb/TU_Darmstadt_Gelbild_Restriktionsverdau_xylBC.png" src="https://static.igem.org/mediawiki/2015/e/eb/ | + | <img class="transparent" alt="https://static.igem.org/mediawiki/2015/e/eb/TU_Darmstadt_Gelbild_Restriktionsverdau_xylBC.png" src="https://static.igem.org/mediawiki/2015/e/eb/TU_Darmstadt_Gelbild_Restriktionsverdau_ xylBC.png"><b>Figure 1</b> Restriction digest of B0034-<i>xylB</i>-B0034-<i>xylC</i> (along with others). You can see the <i>xylB-xylC</i> string (below 2 kbp) and the pSB1C3 backbone (above 2 kbp). DNA marker: 2-Log DNA Ladder (NEB).</p>.</figcaption> |
</figure> | </figure> | ||
| Line 12: | Line 12: | ||
The <a href=" http://parts.igem.org/Part:BBa_K1602000" title="Opens internal link in current window" class="internal link">BBa_K1602000</a> part and the <a href=" http://parts.igem.org/Part:BBa_K1602001" title="Opens internal link in current window" class="internal link">BBa_K1602001</a> part were combined.</p> | The <a href=" http://parts.igem.org/Part:BBa_K1602000" title="Opens internal link in current window" class="internal link">BBa_K1602000</a> part and the <a href=" http://parts.igem.org/Part:BBa_K1602001" title="Opens internal link in current window" class="internal link">BBa_K1602001</a> part were combined.</p> | ||
<p>Firstly the B0034-<em>xylB</em> composite was digested with <em>EcoR</em>I and <em>Spe</em>I while the B0034-<em>xylC</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 restriction digest with <em>EcoR</em>I and <em>Pst</em>I. Plasmid DNA was isolated from positive clones and verified by sanger sequencing.</p> | <p>Firstly the B0034-<em>xylB</em> composite was digested with <em>EcoR</em>I and <em>Spe</em>I while the B0034-<em>xylC</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 restriction digest with <em>EcoR</em>I and <em>Pst</em>I. Plasmid DNA was isolated from positive clones and verified by sanger sequencing.</p> | ||
| + | |||
| + | 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 restriction digest with EcoRI and PstI. Plasmid DNA was isolated from positive clones and verified by sanger sequencing. | ||
</div></html> | </div></html> | ||
Revision as of 16:25, 18 September 2015
K1602056 - B0034-D-Xylonic Acid Dehydratase and B0034-Aldehyde Reductase (B0034-yjhG-B0034-yqhD)
Figure 1 Restriction digest of B0034-xylB-B0034-xylC (along with others). You can see the xylB-xylC string (below 2 kbp) and the pSB1C3 backbone (above 2 kbp). DNA marker: 2-Log DNA Ladder (NEB)..
The BBa_K1602000 part and the BBa_K1602001 part were combined.
Firstly the B0034-xylB composite was digested with EcoRI and SpeI while the B0034-xylC 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 restriction digest with EcoRI and PstI. Plasmid DNA was isolated from positive clones and verified by sanger sequencing.
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 restriction digest with EcoRI and PstI. Plasmid DNA was isolated from positive clones and verified by sanger sequencing.