Team:UNITN-Trento/Notebook/Labposts/06/60

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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" : "fabio-bruno",
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"author" : "emil-gabriele",
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"title" : "bacillus subtilis promoters and plasmids (part 10)!!",
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"title" : "Ligation and transformation of R0010 + Sam sinthethase",
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"content" : "<html> I miniprepped K823025 and obtained two good yields: 293,5 ng/ul and 236,6 ng/ul; I screened both samples (1000 ng digested in 50 ul) and the gel confirmed the presence of the plasmid. We have another one!!Afterwards we reported a summary table of part obtained and their yelds</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Summary table|<html><center><table border=\"1\"><tr><th>Sample</th><th>ng/ul</th></tr><tr><td>K823001 a</td><td>436.8</td></tr><tr><td>b</td><td>275.4</td></tr><tr><td>c</td><td>466.5</td></tr><tr><td>K823002a</td><td>138.0</td></tr><tr><td>b</td><td>134.4</td></tr><tr><td>c</td><td>139.7</td></tr><tr><td>K823003 a</td><td>85.4</td></tr><tr><td>b</td><td>99.9</td></tr><tr><td>c</td><td>118.9</td></tr><tr><td>K143012 a</td><td>178.3</td></tr><tr><td>b</td><td>89.2</td></tr><tr><td>c</td><td>121.4</td></tr><tr><td>K823024 a</td><td>362.1</td></tr><tr><td>d</td><td>209.3</td></tr><tr><td>e</td><td>288.2</td></tr><tr><td>K823026 a</td><td>246.3</td></tr><tr><td>b</td><td>315.8</td></tr><tr><td>c</td><td>550.3</td></tr></table></center></html>}}<html></html>",
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"content" : "<html>Today we added 1&micro;l of SAP to the o/n digestion and then incubated it for 1.5h.Then we performed the ligation of SAMynthetase (previously digested with XbaI and PstI), following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Ligation\">ligation protocol</a>.<br/></html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Ligation mixes|<html><center><table class=\"tn-sp-table\"><tr><td style=\"border:none\"></td><th>Ctrl</th><th>1:1</th><th>1:3</th></tr><tr><td>Buffer</td><td colspan=\"3\">2.5&micro;l</td></tr><tr><td>plasmid</td><td colspan=\"3\">5.73&micro;l</td></tr><tr><td>insert</td><td>0</td><td>3.09&micro;l</td><td>9.27&micro;l</td></tr><tr><td>ligase</td><td colspan=\"3\">1&micro;l</td></tr><tr><td>water</td><td>15.77&micro;l</td><td>12.68&micro;l</td><td>6.50&micro;l</td></tr></table></center></html>}}<html>Afterwards, Gabriele purified the product following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Promega-PCR-Gel\">usual purification protocol</a> and then transformed it in NEB10&beta; cells following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Competent-cells-transformation\">transformation protocol</a>.<br/><br/>Gabriele prepared also 4 inocula from the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Notebook#tn-post-2013-06-18-gabriele-emil\">pSB1C3+R0010 CM-plate</a>, just to be sure that it is (not) there.</html>",
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"tags" : "plasmidPxil-plasmidPspac - plasmidluxABCDE-Pveg-PliaG-PlepA-Plial"
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"tags" : "Plac-SAMsynthetase"
}
}

Latest revision as of 07:57, 3 October 2013

{ "date" : "2013-06-26", "author" : "emil-gabriele", "title" : "Ligation and transformation of R0010 + Sam sinthethase", "content" : "Today we added 1µl of SAP to the o/n digestion and then incubated it for 1.5h.Then we performed the ligation of SAMynthetase (previously digested with XbaI and PstI), following the ligation protocol.

Ligation mixes
Ctrl1:11:3
Buffer2.5µl
plasmid5.73µl
insert03.09µl9.27µl
ligase1µl
water15.77µl12.68µl6.50µl
Afterwards, Gabriele purified the product following the usual purification protocol and then transformed it in NEB10β cells following the transformation protocol.

Gabriele prepared also 4 inocula from the pSB1C3+R0010 CM-plate, just to be sure that it is (not) there.", "tags" : "Plac-SAMsynthetase" }

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