Team:UNITN-Trento/Notebook/Labposts/06/59

From 2013.igem.org

(Difference between revisions)
 
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{
{
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"date" : "2013-06-27",
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"date" : "2013-06-26",
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"author" : "gabriele-emil",
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"author" : "fabio-bruno",
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"title" : "SAMsynthetase and R0010, the neverending story",
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"title" : "bacillus subtilis promoters and plasmids (part 9)!!",
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"content" : "<html>First of all Gabriele quantified the four pSB1C3+R0010 inocula of yesterday.</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Quantification|<html><center><table class=\"tn-sp-table\"><tr><th>Sample</th><th>Quantity (ng/&micro;l)</th></tr><tr><td>#A</td><td>243.7</td></tr><tr><td>#B</td><td>194.1</td></tr><tr><td>#C</td><td>245.3</td></tr><tr><td>#D</td><td>252.9</td></tr></table></center></html>}}<html><br/>Then, the four quantified samples were screened after a digestion with ExoRI-HF and PstI-HF, with the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Digestion\">usual screening protocol</a> (incubated only for 45min, since both the enzymes are HF).<br/><br/>The digestion products were then run on a gel with a transparent loading dye (30% glycerol): each loaded sample contained 16&micro;l of the sample and 4&micro;l of transparent loading dye. Both a 1kb normal ladder and a 100bp transparent ladder were loaded, also the first well was loaded with the usual loading dye.</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Gel|<html><center><table class=\"tn-sp-table\"><tr><th>Loading scheme</th></tr><tr><td><i>Loading dye</i></td></tr><tr><td><i>Empty</i></td></tr><tr><td>100bp ladder</td></tr><tr><td>#A</td></tr><tr><td>#B</td></tr><tr><td>#C</td></tr><tr><td>#D</td></tr><tr><td><i>Empty</i></td></tr><tr><td><i>Empty</i></td></tr><tr><td>1kb ladder</td></tr></table><img src=\"https://static.igem.org/mediawiki/2013/6/62/Tn-20130627-GG_R0010pSB1C3_FAILED.jpg\" width=\"450px\" /><br/><img src=\"https://static.igem.org/mediawiki/2013/0/0b/Tn-20130627-GG_R0010pSB1C3_BIS.JPG\" height=\"450px\" /></center></html>}}<html><br/>Finally, Emil prepared the inocula of the product of ligation of R0010 and SAMsynthetase (in pSB1A2: 6 inocula of the 1:1 ligation product and 3 of the 1:3 ligation producta).</html>",
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"content" : "<html> in order to verify weather the plasmids have been inverted during miniprep purification protocol, or screening protocol or even before, during plating, we made the minipreps out of the remaining inocula and then screened them: the risults were the same, so probably we made a mistake during plating! Now everything is more clear!!In the afternoon we made the inoculum of the last plasmid ( K823025, sent right from the Part’s Registry Headquarters).</html>",
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"tags" : "SAMsynthetase-Plac"
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"tags" : " plasmidPxil-plasmidPspac - plasmidluxABCDE "
}
}

Latest revision as of 07:57, 3 October 2013

{ "date" : "2013-06-26", "author" : "fabio-bruno", "title" : "bacillus subtilis promoters and plasmids (part 9)!!", "content" : " in order to verify weather the plasmids have been inverted during miniprep purification protocol, or screening protocol or even before, during plating, we made the minipreps out of the remaining inocula and then screened them: the risults were the same, so probably we made a mistake during plating! Now everything is more clear!!In the afternoon we made the inoculum of the last plasmid ( K823025, sent right from the Part’s Registry Headquarters).", "tags" : " plasmidPxil-plasmidPspac - plasmidluxABCDE " }