Difference between revisions of "Team:Freiburg/Project/pRIG15 8"
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To insert the sequence for HSV1 glycoprotein G into pSB1C3 we designed Gibson primers with compatible overhangs that also included the start codon ATG. This fragment was amplified via PCR (Link zum Labjournal-Eintrag) and then assembled with the digested pSB1C3 backbone using Gibson assembly. | To insert the sequence for HSV1 glycoprotein G into pSB1C3 we designed Gibson primers with compatible overhangs that also included the start codon ATG. This fragment was amplified via PCR (Link zum Labjournal-Eintrag) and then assembled with the digested pSB1C3 backbone using Gibson assembly. | ||
To prove correct insertion of our fragment we did a test digest (Link Labjournal) and sent the whole plasmid for sequencing. | To prove correct insertion of our fragment we did a test digest (Link Labjournal) and sent the whole plasmid for sequencing. | ||
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+ | Link to genebank file: <a class="media" href="https://static.igem.org/mediawiki/2015/2/22/Freiburg_2015_BBa_K1621002.gb" title="2015_Freiburg_BBa_K1621002" src="https://static.igem.org/mediawiki/2015/2/22/Freiburg_2015_BBa_K1621002.gb">BBa_K1621002.gb</a>. | ||
</p> | </p> | ||
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Revision as of 12:30, 7 September 2015
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pRIG15_8
Infection with the Herpes Simplex Virus 1 leads to life-long persistence of the virus inside the body as a latent infection. 1) The glycoprotein G is one of 11 glycoproteins expressed by HSV1. 2) The sequence we used for our experiment was obtained from Jaaskelainen et al., 2009. 3)
To insert the sequence for HSV1 glycoprotein G into pSB1C3 we designed Gibson primers with compatible overhangs that also included the start codon ATG. This fragment was amplified via PCR (Link zum Labjournal-Eintrag) and then assembled with the digested pSB1C3 backbone using Gibson assembly. To prove correct insertion of our fragment we did a test digest (Link Labjournal) and sent the whole plasmid for sequencing. Link to genebank file: BBa_K1621002.gb.