Team:UNITN-Trento/Notebook/Labposts/07/39

From 2013.igem.org

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{"date" : "2013-07-18","author" : "thomas-michele","title" : "Back to work","content" : "<html>Today, after a long and hard fight with our thesis, we came back to work. In particular we teased the 2012 Registry Distribution and we extracted two new parts: BBa_K537055 (679 bp) and BBa_K537056 (721 bp). These two parts contained two reportes gene, mCherry and Venus. We did this because we wanted to fuse at the C-term of BBa_K1065001 a reporter in order to measure the kinetic of the protein sythesis. A new chapter to add to our team!  </html>","tags" : "EFE-mCherry-Venus"}
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"date" : "2013-07-20",
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"author" : "thomas",
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"title" : "Digestion of AraC-pBAD + EFE and Venus PCR product",
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"content" : "<html>This is the part two of <a href=\"https://2013.igem.org/wiki/index.php?title=Team:UNITN-Trento/Notebook#tn-post-2013-07-18-thomas-michele\">this experiment</a>.Since both Venus (BBa_K537006) and AraC-pBAD + EFE were in pSB1C3 vector, I had to do a PCR on Venus using primers Prefix Fwd and Suffing Rev following <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#pSB1C3-linearization-PCR\">this protocol</a>. The PCR product was then confirmed by an electrophoresis analysis (Venus is 729 bp and KAPA universal Ladder was adopted). <br/></html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Gel Image|<html><center><img src=\"https://static.igem.org/mediawiki/2013/f/fe/Tn-2013_VenusPCR_gel.jpg\" style =\"width: 450px\"></center></html>}}<html><br/>The PCR product was then purified and quantified at the NanoDrop.Once I obtained Venus without his backbone, I proceeded digesting all the PRC Venus product with NgoMIV and PstI and 2-3 ug of AraC-pBAD + EFE with AgeI and PstI following <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Digestion\">the PCR product digestion protocol</a>. The digestion products were finally purified, quantified and stored at -20&deg;C.<br/> </html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Quantification Results|<html><center><table class=\"tn-sp-table\"><tr><td>AraC-pBAD + EFE</td><td>33.5 ng/ul</td></tr><tr><td>Venus PCR product</td><td>22.8 ng/ul</td></tr></table></center></html>}}<html></html>",
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"tags" : "EFE-Venus"
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}
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Latest revision as of 10:41, 3 October 2013

{"date" : "2013-07-18","author" : "thomas-michele","title" : "Back to work","content" : "Today, after a long and hard fight with our thesis, we came back to work. In particular we teased the 2012 Registry Distribution and we extracted two new parts: BBa_K537055 (679 bp) and BBa_K537056 (721 bp). These two parts contained two reportes gene, mCherry and Venus. We did this because we wanted to fuse at the C-term of BBa_K1065001 a reporter in order to measure the kinetic of the protein sythesis. A new chapter to add to our team! ","tags" : "EFE-mCherry-Venus"}