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

From 2013.igem.org

(Difference between revisions)

Latest revision as of 10:33, 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" : "2013-07-02","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" : "<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" : "E0840-K823026-K823024"}
-	"date" : "2013-07-03",
-	"author" : "gabriele",
-	"title" : "An easy day...",
-	"content" : "<html>First of all I counted the colonies from <a href=\"https://2013.igem.org/wiki/index.php?title=Team:UNITN-Trento/Notebook#tn-post-2013-07-02-gabriele\">yesterday</a>:<br/><br/><center><table class=\"tn-sp-table\"><tr><th>plate</th><th>#colonies</th></tr><tr><td>Ctrl</td><td>4</td></tr><tr><td>0.5:1</td><td>2</td></tr><tr><td>0.5:2</td><td>0</td></tr></table></center><br/><hr><br/>Then, I tried again to linearize pSB1C3 (this time, starting from the circular), but I failed.</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Gel|<html><center><table class=\"tn-sp-table\"><tr><th colspan=\"4\">Loading scheme</th></tr><tr><td>1kb ladder</td><td>G1</td><td>G2</td><td>G3</td></tr></table><img src=\"https://static.igem.org/mediawiki/2013/d/d3/Tn-20130703-GGCM_linearizationpSB1C3_FAILED.jpg\" alt=\"Gel\" width=\"450px\"></center></html>}}<html></html>",
-	"tags" : "SAMsynthetase-pSB1C3"
-}