Team:UNITN-Trento/Notebook/Labposts/06/66

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(Difference between revisions)

Latest revision as of 08:00, 3 October 2013

{ "date" : "2013-06-29", "author" : "emil", "title" : "Purification and screening of the GFPs E0840 and E0240", "content" : " I did the miniprep of the 8 inocula(2 GFP constructs) following the vacuum protocol then I have quantified the products with the following results:

Quantification

Sample	Quantification
BBa_E0840/1	484.6ng/µl
BBa_E0840/2	376ng/µl
BBa_E0840/3	481.8ng/µl
BBa_E0840/4	475ng/µl
BBa_E0240/1	361ng/µl
BBa_E0240/2	454.4ng/µl
BBa_E0240/3	368ng/µl
BBa_E0240/4	365.8ng/µl

Then I did the screening of the samples E0840/1,E0840/3,E0240/2,E0240/4. I digested 800 ng of each sample with EcoR1-HF and Pst1-HF following the digestion protocol for screening. After 40 minutes I loaded the samples on a gel with the following results:

Gel order

Sample	Well
Ladder 1kb Fermentas	1
BBa_E0840/1	2
BBa_E0840/3	3
BBa_E0240/2	4
BBa_E0240/4	5

image

We can see 2 bands for well: the upper is the backbone(pSB1A2 2079 bp), the lower is presumably the GFP(BBa_E0240 826 bp,BBa_E0840 828 bp)(pefix and suffix escluded ?).", "tags" : "E0840-E0240" }

@@ Line 1: / Line 1: @@
 {
-	"date" : "2013-06-10",
+	"date" : "2013-06-29",
-	"author" : "caterina-fabio",
+	"author" : "emil",
-	"title" : "The Registry 2013 is deflowered!",
+	"title" : "Purification and screening of the GFPs E0840 and E0240",
-	"content" : "We extracted <b>BBa_K143012</b>, <b>BBa_K823000</b>, <b>BBa_K823002</b> and <b>BBa_K823003</b> and then transformed them into NEB10&beta; cells.",
+	"content" : "<html> I did the miniprep of  the 8 inocula(2 GFP constructs) following the <a href=\"http://ita.promega.com/~/media/Files/Resources/ProtCards/Wizard%20Plus%20SV%20Minipreps%20DNA%20Purification%20System%20Quick%20Protocol.pdf\">vacuum protocol</a> then I have quantified the products with the following results:</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Quantification|<html><center><table class=\"tn-sp-table\"><tr><th>Sample</th><th>Quantification</th></tr><tr><td>BBa_E0840/1</td><td> 484.6ng/&micro;l</td></tr><tr><td>BBa_E0840/2</td><td> 376ng/&micro;l</td></tr><tr><td>BBa_E0840/3</td><td> 481.8ng/&micro;l</td></tr><tr><td>BBa_E0840/4</td><td> 475ng/&micro;l</td></tr><tr><th>BBa_E0240/1</th><th> 361ng/&micro;l</th></tr><tr><td>BBa_E0240/2</td><td> 454.4ng/&micro;l</td></tr><tr><td>BBa_E0240/3</td><td> 368ng/&micro;l</td></tr><tr><td>BBa_E0240/4</td><td> 365.8ng/&micro;l</td></tr></table></center></html>}}<html>Then I did the screening of the samples E0840/1,E0840/3,E0240/2,E0240/4. I digested 800 ng of each sample with EcoR1-HF and Pst1-HF following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Digestion\">digestion protocol for screening</a>. After 40 minutes I loaded the samples on a gel with the following results:</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Gel order|<html><center><table class=\"tn-sp-table\"><tr><th>Sample</th><th>Well</th></tr><tr><td>Ladder 1kb Fermentas</td><td>1</td></tr><tr><td>BBa_E0840/1</td><td>2</td></tr><tr><td>BBa_E0840/3</td><td>3</td></tr><tr><td>BBa_E0240/2</td><td>4</td></tr><tr><td>BBa_E0240/4</td><td>5</td></tr></table></center></html>}}{{:Team:UNITN-Trento/Templates/Styles/Spoiler|image|<html> <img src=\"https://static.igem.org/mediawiki/2013/4/4b/Tn-20130629-Et_GFP.jpg\" width=\"450px\" /></html>}}<html> We can see 2 bands for well: the upper is the backbone(pSB1A2 2079 bp), the lower is presumably the GFP(BBa_E0240 826 bp,BBa_E0840 828 bp)(pefix and suffix escluded ?).</html>",
-	"tags" : "Pveg-PliaG-PlepA"
+	"tags" : "E0840-E0240"
 }