Difference between revisions of "Team:TU Darmstadt/Notebook/sec1/K1602012"
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− | <h3> | + | <h3>K1602012 - <small>D</small>-Xylonic acid dehydratases yjhG</h3> |
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<figure class="rightFig" style="width:30%"> | <figure class="rightFig" style="width:30%"> | ||
− | <img src="https://static.igem.org/mediawiki/2015/8/86/ | + | <img src="https://static.igem.org/mediawiki/2015/8/86/Agar_gel_c3_ cadA.png" width=100% height=100%> |
<figcaption><br><b>Figure 1</b> PCR of cadA (X). The size of the amplified product was around 1.5 kbp. DNA marker: 2-Log DNA Ladder (NEB).</figcaption> | <figcaption><br><b>Figure 1</b> PCR of cadA (X). The size of the amplified product was around 1.5 kbp. DNA marker: 2-Log DNA Ladder (NEB).</figcaption> | ||
</figure> | </figure> | ||
− | The < | + | The <i>yjhG</i>-gene was isolated from K12 E.coli and amplified through colony PCR using the <i>yjhG</i> (FW) and <i>yjhG</i> (REV) oligonucleotides. The PCR product was verified by agarose gel electrophoresis, digested with Dpn1 for 120 minutes and subsequently purified.The <i>YjhG</i> amplicon was cut with EcoRI and PstI and ligated into a pSB1C3 vector using T4-ligase. <i>E.Coli</i> Top10 has been transformed with the ligation product by heat shock transformation and the cells were spread out on a CMP agar plate. Clones were screened with colony PCR using VF2 and VR oligonucleotides. Plasmid DNA was isolated from positive clones which were verified by sanger sequencing. |
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Revision as of 15:05, 17 September 2015
K1602012 - D-Xylonic acid dehydratases yjhG
![](https://static.igem.org/mediawiki/2015/8/86/Agar_gel_c3_ cadA.png)
Figure 1 PCR of cadA (X). The size of the amplified product was around 1.5 kbp. DNA marker: 2-Log DNA Ladder (NEB).