Team:UNITN-Trento/Notebook/Labposts/07/06

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Revision as of 08:22, 3 October 2013

{ "date" : "2013-07-02", "author" : "emil", "title" : "Purification of the digestion products (K823026, K823024, E0840) ligation of the GFP in the 2 backbones and transformation of the ligation products ", "content" : "This morning I added 1µl of DPN1 to the GFP ligation and 1µl of SAP(alkaline phosphatase) to the 2 backbones.After 1h 30 min I have stopped the reaction by putting the reaction tubes at 80 C°.Afterwards I have purified the products following the purification protocol.Then I quantified the products with the following results:

Quantification

Sample	Quantification
BBa_E0840	15.8ng/µl
BBa_K823026	37.6ng/µl
BBa_K823024	36.4ng/µl

Then I performed the ligation(1:1,1:3,CTRL) of the GFP with the 2 backbone individually and following the ligation protocol.Finally I transformed 10µl of the ligation protocol in Neb10β and plate them on ampicillin LB agar following the transformation protocol.", "tags" : "E0840-K823026-K823024" }

@@ Line 1: / Line 1: @@
 {
-	"date" : "2013-07-22",
+	"date" : "2013-07-02",
 	"author" : "emil",
-	"title" : "The incompetent Bacillus subtilis saga: The mistaken antibiotic ",
+	"title" : "Purification of the digestion products (K823026, K823024, E0840) ligation of the GFP in the 2 backbones and transformation of the ligation products ",
-	"content" : "<html>Unfortunately the previous attempt to transform failed maybe due to the mistaken concentration of antibiotic so I restarted generating a stock solution of Kanamicin(10mg/ml).I did an inocula in LB of Bacillus overnight from the stock solution with 50% of glicerol.</html>",
+	"content" : "<html>This morning I added 1&micro;l of DPN1 to the GFP ligation and 1&micro;l of SAP(alkaline phosphatase) to the 2 backbones.After 1h 30 min I have stopped the reaction by putting the reaction tubes at 80 C&deg;.Afterwards I have purified the products following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Promega-PCR-Gel\"> purification protocol</a>.Then I quantified the products with the following results:</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Quantification|<html><table>     <tr><th>Sample</th><th>Quantification</th></tr><tr><td>BBa_E0840</td><td> 15.8ng/&micro;l</td></tr><tr><td>BBa_K823026</td><td> 37.6ng/&micro;l</td></tr><tr><td>BBa_K823024</td><td> 36.4ng/&micro;l</td></tr></table></html>}}<html>Then I performed the ligation(1:1,1:3,CTRL) of the GFP with the 2 backbone individually and following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Ligation\"> ligation protocol</a>.Finally I transformed 10&micro;l of the ligation protocol in Neb10&beta; and plate them on ampicillin LB agar following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Competent-cells-transformation\"> transformation protocol</a>.</html>",
-	"tags" : "Bacillus subtilis"
+	"tags" : "E0840-K823026-K823024"
 }