Team:UNITN-Trento/Notebook/Labposts/06/07

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{
{
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"date" : "2013-06-11",
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"date" : "2013-06-07",
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"author" : "emil-fabio",
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"author" : "viola-fabio-emil",
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"title" : "",
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"title" : "Purification and PCR of pSB1C3 linearized, gel",
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"content" : "<html><h5>Inoculum of three B.subtilis backbone</h5>We have done the inoculum of three Bacillus-specific backbone (previously transformed in NEB and plated on Ampicillin LB Agar) in 19 50ml Falcon- 7 with the part BBa_K823023- 6 with the part BBa_K823022- 6 with the part BBa_K823027The Falcons were put in the incubator at 37 °C o/n.<h5>Digestion of SAMsynthetase and psB1C3</h5>We have digested the SAMsynthetase gene (previously amplificated and purificated)and the linerarized vector psB1C3 with the enzyme Xba1 and Pst1 exploiting the same protocol. Then we have incubated the mix at 37C o/n.</html>",
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"content" : "<html>We purified with the Quick Reference purification kit(GE) and made a PCR of the linearized plasmid pSB1C3 using the prefix and suffix primers  and the Phusion polimerase following <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Phusion-PCR\">its protocol</a>. Then we took the PCR samples and checked the results with an electrophoresis gel  using the ladder Generuler 1 kb Plus DNA Ladder of Fermentas:</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Gel Image|<html><img id=\"post_img\" src=\" https://static.igem.org/mediawiki/2013/8/83/Tn-20130607-pSB1C3_linear.png\" width=\"550px\"/></html>}}<html>As we can see the plasmid, stopped at 2 kb ca (pSB1C3=2070kb).</html>",
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"tags" : "SAMsynthetase-pSBBs1C-pSBBs4S-pSBBs2E"
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"tags" : "pSB1C3"
}
}

Revision as of 07:40, 3 October 2013

{ "date" : "2013-06-07", "author" : "viola-fabio-emil", "title" : "Purification and PCR of pSB1C3 linearized, gel", "content" : "We purified with the Quick Reference purification kit(GE) and made a PCR of the linearized plasmid pSB1C3 using the prefix and suffix primers and the Phusion polimerase following its protocol. Then we took the PCR samples and checked the results with an electrophoresis gel using the ladder Generuler 1 kb Plus DNA Ladder of Fermentas:

Gel Image
As we can see the plasmid, stopped at 2 kb ca (pSB1C3=2070kb).", "tags" : "pSB1C3" }

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