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 | + | 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 <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Labjournals/Cloning/August#PCR6-18" title="PCR6-18">PCR</a> and then assembled with the digested pSB1C3 backbone using Gibson assembly. |
| − | To prove correct insertion of our fragment we did a test digest | + | To prove correct insertion of our fragment we did a <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Labjournals/Cloning/August#TD8_10_18" title="TD8_10_18">test digest</a> and sent the whole plasmid for sequencing. |
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Revision as of 14:41, 7 September 2015
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 and then assembled with the digested pSB1C3 backbone using Gibson assembly. To prove correct insertion of our fragment we did a test digest and sent the whole plasmid for sequencing. Link to genebank file: BBa_K1621002.gb.
