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

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(Difference between revisions)
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{
{
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"date" : "2013-07-04",
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"date" : "2013-07-20",
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"author" : "fabio-viola",
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"author" : "thomas",
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"title" : "BACILLUS SUBTILIS COMES BACK TO LIFE!",
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"title" : "Digestion of AraC-pBAD + EFE and Venus PCR product",
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"content" : "<html>  today we made glycerol stock tubes from the remaining cultures and from the inocula made yesterday, but before that we collected some bacteria to make other two liquid cultures just to have more glycerol stock tubes to put at -80°</html>",
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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" : " Bacillus subtilis "
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"tags" : "EFE-Venus"
}
}

Revision as of 08:35, 3 October 2013

{ "date" : "2013-07-20", "author" : "thomas", "title" : "Digestion of AraC-pBAD + EFE and Venus PCR product", "content" : "This is the part two of this experiment.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 this protocol. The PCR product was then confirmed by an electrophoresis analysis (Venus is 729 bp and KAPA universal Ladder was adopted).

Gel Image

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 the PCR product digestion protocol. The digestion products were finally purified, quantified and stored at -20°C.
Quantification Results
AraC-pBAD + EFE33.5 ng/ul
Venus PCR product22.8 ng/ul
", "tags" : "EFE-Venus" }

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