Team:UNITN-Trento/Notebook/Labposts/06/20

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(Difference between revisions)

Latest revision as of 07:44, 3 October 2013

{ "date" : "2013-06-12", "author" : "Gabriele", "title" : "Ligation and transformation", "content" : "Ligation of SAMsynthetase and pSB1C3 with and without RFP, so I prepared four samples:
1) pSB1C3 with RFP (Ctrl_1)
2) pSB1C3 with RFP + SAMsynthetase (1:2)
3) pSB1C3 without RFP (Ctrl_2)
4) pSB1C3 without RFP (1:2)
The samples were prepared and ligation was performed following the ligation protocol.

Ligation parameters

Plasmid concentration: 27.8ng/µl
Plasmid length: 2070bp
Insert concentration: 13.2ng/µl
Insert length: 1155bp
Used plasmid: 250µl
Volume of reaction: 35µl
Buffer concentration: 10X

Transformation in NEB10β cells was performed following the transformation protocol.Plates in incubator ON at 37°C static, more than 16 hours.", "tags" : "SAMsynthetase" }

@@ Line 1: / Line 1: @@
 {
-	"date" : "2013-06-13",
+	"date" : "2013-06-12",
-	"author" : "gabriele",
+	"author" : "Gabriele",
-	"title" : "Ligation results: let's screen it!",
+	"title" : "Ligation and transformation",
-	"content" : "<html>Both the 1:2 ligation plates have number of colonies comparable with their control:<br/><ul><li>SAMsynthetase+RFP (1:2) = too many to count.</li><li>SAMsynthetase+RFP (ctrl) = too many to count.</li><li>SAMsynthetase (1:2) = 11 colonies.</li><li>SAMsynthetase (ctrl) = 12 colonies.</li></ul><br/></html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|SAMsynthetase+pSB1C3[RFP] (1:2) plate|<html><center><img src=\"https://static.igem.org/mediawiki/2013/8/8e/Tn-20130613-SAM_RFP_1_2.JPG\" width=\"300px\" /></center></html>}}<html></html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|SAMsynthetase+pSB1C3[RFP] (ctrl) plate|<html><center><img src=\"https://static.igem.org/mediawiki/2013/c/ce/Tn-20130613-SAM_RFP_ctrl.JPG\" width=\"300px\" /></center></html>}}<html></html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|SAMsynthetase+pSB1C3 (1:2) plate|<html><center><img src=\"https://static.igem.org/mediawiki/2013/f/f9/Tn-20130613-SAM_1_2.JPG\" width=\"300px\" /></center></html>}}<html></html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|SAMsynthetase+pSB1C3 (ctrl) plate|<html><center><img src=\"https://static.igem.org/mediawiki/2013/b/bd/Tn-20130613-SAM_ctrl.JPG\" width=\"300px\" /></center></html>}}<html><br/>Performed 5 inocula for each of the two 1:2 plates, in 5ml LB with 5&micro;l CM. Tomorrow miniprep, quantification, digestion and gel to control what happened.</html>",
+	"content" : "<html>Ligation of SAMsynthetase and pSB1C3 with and without RFP, so I prepared four samples:<br/>1) pSB1C3 with RFP (Ctrl_1)<br/>2) pSB1C3 with RFP + SAMsynthetase (1:2)<br/>3) pSB1C3 without RFP (Ctrl_2)<br/>4) pSB1C3 without RFP (1:2)<br/>The samples were prepared and ligation was performed following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Ligation\">ligation protocol</a>.<br/><br/></html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Ligation parameters|<html><ul><li>Plasmid concentration: 27.8ng/&micro;l</li><li>Plasmid length: 2070bp</li><li>Insert concentration: 13.2ng/&micro;l</li><li>Insert length: 1155bp</li><li>Used plasmid: 250&micro;l</li><li>Volume of reaction: 35&micro;l</li><li>Buffer concentration: 10X</li></ul></html>}}<html><br/>Transformation in NEB10&beta; cells was performed following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Competent-cells-transformation\">transformation protocol</a>.Plates in incubator ON at 37&deg;C static, more than 16 hours.</html>",
 	"tags" : "SAMsynthetase"
 }