Team:UNITN-Trento/Notebook/Labposts/06/34

From 2013.igem.org

(Difference between revisions)
(Created page with "{ "date" : "2013-06-18", "author" : "fabio-bruno", "title" : "bacillus subtilis promoters (part3)!!", "content" : "<html> from the previous inoculum ( with NEB10b cells) we c...")
 
(One intermediate revision not shown)
Line 1: Line 1:
{
{
-
"date" : "2013-06-18",
+
"date" : "2013-06-17",
-
"author" : "fabio-bruno",
+
"author" : "thomas",
-
"title" : "bacillus subtilis promoters (part3)!!",
+
"title" : "Cloning of EFE in pSB1C3",
-
"content" : "<html> from the previous inoculum ( with NEB10b cells) we could only take K823000 ( duplicate) and K143012 (triplicate). Then we performed the minipreps and quantified the yields:K823000 a = 30.7 ng/ulK823000 b = 34.2 ng/ulK143012a = 26.8 ng/ulK143012b = 30.3 ng/ulK143012c = 29.5 ng/ulAs far as NEB5a are concerned, from the inoculum we made our cells competent.In the afternoon we transformed these cells with our promoters.</html>",
+
"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.  ",
-
"tags" : "Pveg-PliaG-PlepA"
+
"tags" : "EFE"
}
}

Latest revision as of 07:49, 3 October 2013

{ "date" : "2013-06-17", "author" : "thomas", "title" : "Cloning of EFE in pSB1C3", "content" : "I began the week doing some minipreps (x5 of EFE and x5 of pSB1C3+RFP) following this protocol.

Quantification results
SampleEFEpSB1C3
1253,8 ng/ul161,9 ng/ul
1243,5 ng/ul160,9 ng/ul
3261,4 ng/ul142,5 ng/ul
4218,0 ng/ul168,4 ng/ul
5299,1 ng/ul sample lost
I then digested 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 ligation and transformation in NEB10B cells. ", "tags" : "EFE" }