Team:UNITN-Trento/Notebook/Labposts/06/47

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(Created page with "{ "date" : "2013-06-07", "author" : "gabriele-michele", "title" : "Third SAMsynthetase synthase extraction attempt - TOO SAD", "content" : " We've tried another combination t...")
 
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{
{
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"date" : "2013-06-07",
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"date" : "2013-06-21",
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"author" : "gabriele-michele",
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"author" : "fabio-bruno",
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"title" : "Third SAMsynthetase synthase extraction attempt - TOO SAD",
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"title" : "bacillus subtilis promoters and  plasmids (part 6)!!",
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"content" : " We've tried another combination to maximize the outputs of the PCR done to amplify the SAMsynthetase synthase gene from an extract of E.coli genomic DNA (strain MG1655). This time we used the <I> RBC Taq DNA Polymerase </I> protocol but with a mix of this polymerase (0.25 ul) and the Phusion polymerase (0.3 ul). We've prepared two identical samples to see who is the better PCR maker! :) {{:Team:UNITN-Trento/Templates/Styles/Spoiler|Gel Image|<html><img src=\"https://static.igem.org/mediawiki/2013/c/c2/Tn-20130607-SAMsynth_pcr_gire-pedro.jpg\" /></html>}}As you can see from the picture we both obtained great results",
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"content" : "<html> The last battle!!! Today we screened successfully three parts, K823003, K823002, K143012 . We used a brand new formula: first of all we digested 1500 ng of DNA in 50 ul. Then we changed the gel composition (1,5 % agarose, 3,5 ul of etidium bromide in 35 ml of gel). Finally we loaded the 100 bp marker along with the 1000kb marker and the parts with a loading dye missing of the dye ( 30% glicerole solution instead). We put this unusual gel in the electrophoresis chamber for 20 minutes. We obtained dim but still present traces.In the afternoon we extracted a new Bacillus promoter (PliaI, K823001) from 2013 registry kit (plate 1 20c) and transformed both in 100 ul of NEB10b and NEB5a.We also did the inocula for the parts plated yesterday.</html>",
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"tags" : "SAMsynthetase"
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"tags" : " Pveg-PliaG-PlepA-plasmidPxil-plasmidPspac-Plial"
}
}

Latest revision as of 07:53, 3 October 2013

{ "date" : "2013-06-21", "author" : "fabio-bruno", "title" : "bacillus subtilis promoters and plasmids (part 6)!!", "content" : " The last battle!!! Today we screened successfully three parts, K823003, K823002, K143012 . We used a brand new formula: first of all we digested 1500 ng of DNA in 50 ul. Then we changed the gel composition (1,5 % agarose, 3,5 ul of etidium bromide in 35 ml of gel). Finally we loaded the 100 bp marker along with the 1000kb marker and the parts with a loading dye missing of the dye ( 30% glicerole solution instead). We put this unusual gel in the electrophoresis chamber for 20 minutes. We obtained dim but still present traces.In the afternoon we extracted a new Bacillus promoter (PliaI, K823001) from 2013 registry kit (plate 1 20c) and transformed both in 100 ul of NEB10b and NEB5a.We also did the inocula for the parts plated yesterday.", "tags" : " Pveg-PliaG-PlepA-plasmidPxil-plasmidPspac-Plial" }

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