Difference between revisions of "Part:BBa K4585009"
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The objective is to perform homologous recombination with the Degron homologous recombination fragment , The synthetic pcDNA3.1 (+) -3 HA-Gal 4-VP64-Degron-NLS plasmid was used for subsequent experiments. | The objective is to perform homologous recombination with the Degron homologous recombination fragment , The synthetic pcDNA3.1 (+) -3 HA-Gal 4-VP64-Degron-NLS plasmid was used for subsequent experiments. | ||
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| + | <h2 class="pageContent-main__title"> | ||
| + | <!--put tile here, <h2>title</h2>, class="pageContent-main__title" means it is the main title--> | ||
| + | pcDNA3.1(+)-3×HA-Gal4-VP64-Degron-NLS linearized vector | ||
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| − | < | + | <p>We based on the sequence of the pcDNA3.1(+)-3×HA-Gal4-VP64-NLS plasmid. Special primers were designed to cut the plasmid into a linear sequence of length of about 6237bp by PCR. The two ends of this linear sequence can be complementary to the Degron homologous recombination fragment of the homologous Degron fragment for homologous recombination to obtain the target product pcDNA3.1(+)-3×HA-Gal4-VP64-Degron-NLS plasmid. |
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| + | <!--put text here, <p>content</p>--> | ||
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| + | <h2 class="pageContent-main__title pageContent-main__subtitle"> | ||
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| + | 1 Pattern Diagram | ||
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| + | <p style="text-align: center;"> | ||
| + | <img width="400px" src="https://static.igem.wiki/teams/4585/wiki/pcdna3-1-3-ha-gal4-vp64-degron-nls-linearized-vector/pcdna3-1-3-ha-gal4-vp64-degron-nls-linearized-vector.png"></p> | ||
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| + | <p style="width: 80%;text-align:center;font-size: .9rem; margin: -1rem auto 1rem auto; color: #888;">Fig.1 The agarose gel electrophoresis of pcDNA3.1(+)-3×HA-Gal4-VP64-Degron-NLS linearized vector</p> | ||
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| + | <h2 class="pageContent-main__title pageContent-main__subtitle"> | ||
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| + | 2 Experiment | ||
| + | </h2> | ||
| + | <h2 class="pageContent-main__title pageContent-main__subtitle"> | ||
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| + | 2.1 Method | ||
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| + | <div class="pageContent-main__textBox"> | ||
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| + | <p>Based on the designed pcDNA3.1 (+) -3 HA-Gal 4-VP64-NLS plasmid , Design of the new pcDNA3.1 (+) -3 HA-Gal 4-VP64-Degron-NLS linearized vectror | ||
| + | </p> | ||
| + | <!--put text here, <p>content</p>--> | ||
| + | </div> | ||
| + | <h2 class="pageContent-main__title pageContent-main__subtitle"> | ||
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| + | 2.2 Results | ||
| + | </h2> | ||
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| + | <!--all the content must included in this div--> | ||
| + | <p style="text-align: center;"> | ||
| + | <img width="400px" src="https://static.igem.wiki/teams/4585/wiki/the-model-diagram-of-pcdna3-1-3-ha-gal4-vp64-degron-nls111.png"></p> | ||
| + | <br /> | ||
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| + | <p style="width: 80%;text-align:center;font-size: .9rem; margin: -1rem auto 1rem auto; color: #888;">Fig.1 The model diagram of pcDNA3.1(+)-3×HA-GAL4-VP64-Degron-NLS</p> | ||
| + | </div> | ||
| + | <h2 class="pageContent-main__title pageContent-main__subtitle"> | ||
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| + | 3.Caution | ||
| + | </h2> | ||
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| + | <p>The two ends of the pcDNA3.1(+)-3×HA-Gal4-VP64-Degron-NLS linearized vectror need to be complementary to the two ends of the corresponding recombinant fragments to ensure the success of homologous recombination ligation. | ||
| + | </p> | ||
| + | </div> | ||
| + | |||
| + | </body> | ||
| + | </html> | ||
Revision as of 11:32, 9 October 2023
pcDNA3.1(+)-3xHA-Gal4-VP64-Degron-NLS linearized vector
The objective is to perform homologous recombination with the Degron homologous recombination fragment , The synthetic pcDNA3.1 (+) -3 HA-Gal 4-VP64-Degron-NLS plasmid was used for subsequent experiments.
pcDNA3.1(+)-3×HA-Gal4-VP64-Degron-NLS linearized vector
We based on the sequence of the pcDNA3.1(+)-3×HA-Gal4-VP64-NLS plasmid. Special primers were designed to cut the plasmid into a linear sequence of length of about 6237bp by PCR. The two ends of this linear sequence can be complementary to the Degron homologous recombination fragment of the homologous Degron fragment for homologous recombination to obtain the target product pcDNA3.1(+)-3×HA-Gal4-VP64-Degron-NLS plasmid.
1 Pattern Diagram
Fig.1 The agarose gel electrophoresis of pcDNA3.1(+)-3×HA-Gal4-VP64-Degron-NLS linearized vector
2 Experiment
2.1 Method
Based on the designed pcDNA3.1 (+) -3 HA-Gal 4-VP64-NLS plasmid , Design of the new pcDNA3.1 (+) -3 HA-Gal 4-VP64-Degron-NLS linearized vectror
2.2 Results
Fig.1 The model diagram of pcDNA3.1(+)-3×HA-GAL4-VP64-Degron-NLS
3.Caution
The two ends of the pcDNA3.1(+)-3×HA-Gal4-VP64-Degron-NLS linearized vectror need to be complementary to the two ends of the corresponding recombinant fragments to ensure the success of homologous recombination ligation.