Team:UNITN-Trento/Notebook/Labposts/06/30

From 2013.igem.org

(Difference between revisions)
 
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{
{
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"date" : "2013-06-17",
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"date" : "2013-06-14",
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"author" : "thomas",
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"author" : "bruno",
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"title" : "Cloning of EFE in pSB1C3",
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"title" : "Transformation efficiency kit results",
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"content" : "I began the week doing some minipreps (x5 of EFE and x5 of pSB1C3+RFP) following <html><a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#miniprep\">this protocol</a></html>. {{:Team:UNITN-Trento/Templates/Styles/Spoiler|Quantification results|<html><center><table class=\"tn-sp-table\"><tr><th>Sample</th><th>EFE</th><th>pSB1C3</th></tr><tr><td>1</td><td>253,8 ng/ul</td><td>161,9 ng/ul</td></tr><tr><td>1</td><td>243,5 ng/ul</td><td>160,9 ng/ul</td></tr><tr><td>3</td><td>261,4 ng/ul</td><td>142,5 ng/ul</td></tr><tr><td>4</td><td>218,0 ng/ul</td><td>168,4 ng/ul</td></tr><tr><td>5</td><td>299,1 ng/ul</td>                <td>sample lost</td></tr></table></center></html>}}I then <html><a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Digestion\">digested</a></html> the linearized pSB1C3 (500 ng, kindly offered by Caterina), pSB1C3 + RFP (500 ng) and EFE (1000 ng) with EcorI and PstI following the 2A assembly protocol.After that, I proceeded with the <html><a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Ligation\">ligation</a></html> and <html><a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Competent-cells-transformation\">transformation</a></html> in NEB10B cells.  ",
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"content" : "<html>The day after transformation I have check the grow of the our cell line and I have find only 4 colony in the more concentrated plate with DNA. This results is probably due to the very low concentration of DNA insert in the cell line (only 50pg of DNA). However, the presence of cell indicates that the efficiency of the cell is sufficient for our experiments.</html>",
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"tags" : "EFE"
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"tags" : "RFP"
}
}

Latest revision as of 07:47, 3 October 2013

{ "date" : "2013-06-14", "author" : "bruno", "title" : "Transformation efficiency kit results", "content" : "The day after transformation I have check the grow of the our cell line and I have find only 4 colony in the more concentrated plate with DNA. This results is probably due to the very low concentration of DNA insert in the cell line (only 50pg of DNA). However, the presence of cell indicates that the efficiency of the cell is sufficient for our experiments.", "tags" : "RFP" }