Team:UNITN-Trento/Notebook/Labposts/06/67

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Latest revision as of 08:00, 3 October 2013

{ "date" : "2013-06-27", "author" : "gabriele-emil", "title" : "SAMsynthetase and R0010, the neverending story", "content" : "First of all Gabriele quantified the four pSB1C3+R0010 inocula of yesterday.

Quantification

Sample	Quantity (ng/µl)
#A	243.7
#B	194.1
#C	245.3
#D	252.9

Then, the four quantified samples were screened after a digestion with ExoRI-HF and PstI-HF, with the usual screening protocol (incubated only for 45min, since both the enzymes are HF).

The digestion products were then run on a gel with a transparent loading dye (30% glycerol): each loaded sample contained 16µl of the sample and 4µl of transparent loading dye. Both a 1kb normal ladder and a 100bp transparent ladder were loaded, also the first well was loaded with the usual loading dye.

Gel

Loading scheme
Loading dye
Empty
100bp ladder
#A
#B
#C
#D
Empty
Empty
1kb ladder

Finally, Emil prepared the inocula of the product of ligation of R0010 and SAMsynthetase (in pSB1A2: 6 inocula of the 1:1 ligation product and 3 of the 1:3 ligation producta).", "tags" : "SAMsynthetase-Plac" }

@@ Line 1: / Line 1: @@
 {
-	"date" : "2013-06-28",
+	"date" : "2013-06-27",
-	"author" : "thomas",
+	"author" : "gabriele-emil",
-	"title" : "Ethylene detection through microGC!!!",
+	"title" : "SAMsynthetase and R0010, the neverending story",
-	"content" : "<html>In order to see if our EFE enzyme is effectively active and functional I made a gas-cromatography analysis.<br/> Starting from an overnight culture, I diluted it 1:100 and I incubated it until it reached 0,5 O.D.600. After that, I added 5mM of Arabinose and I incubated the culture at 37&deg;C for about 4-5 hours. In the I connected the vial previously keeped ermetically closed at the micro GC and I took the measure. I did this work for three samples: negative control (not induced), 5mM Arabinose V=1,5ml and 5mM Arabinose V=3ml.  </html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Results table|<html><center><table class=\"tn-sp-table\"><tr><th>Sample</th><th>Ethylene detected</th></tr><tr><td>Not induced</td><td>0&plusmn;15 ppm </td></tr><tr><td>5mM Arabinose V=1,5ml</td><td>30&plusmn;15 ppm </td></tr><tr><td>5mM Arabinose V=3ml</td><td>70&plusmn;15 ppm </td></tr></table></center></html>}}{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Apparatus image|<html><center><img src=\"https://static.igem.org/mediawiki/2013/8/8c/Tn-20130628-Apparatus.JPG\" style =\"width: 450px\"></center></html>}}<html>As you can see, seems that the device is correct and functionally active! Definetely a good result!</html>",
+	"content" : "<html>First of all Gabriele quantified the four pSB1C3+R0010 inocula of yesterday.</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Quantification|<html><center><table class=\"tn-sp-table\"><tr><th>Sample</th><th>Quantity (ng/&micro;l)</th></tr><tr><td>#A</td><td>243.7</td></tr><tr><td>#B</td><td>194.1</td></tr><tr><td>#C</td><td>245.3</td></tr><tr><td>#D</td><td>252.9</td></tr></table></center></html>}}<html><br/>Then, the four quantified samples were screened after a digestion with ExoRI-HF and PstI-HF, with the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Digestion\">usual screening protocol</a> (incubated only for 45min, since both the enzymes are HF).<br/><br/>The digestion products were then run on a gel with a transparent loading dye (30% glycerol): each loaded sample contained 16&micro;l of the sample and 4&micro;l of transparent loading dye. Both a 1kb normal ladder and a 100bp transparent ladder were loaded, also the first well was loaded with the usual loading dye.</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Gel|<html><center><table class=\"tn-sp-table\"><tr><th>Loading scheme</th></tr><tr><td><i>Loading dye</i></td></tr><tr><td><i>Empty</i></td></tr><tr><td>100bp ladder</td></tr><tr><td>#A</td></tr><tr><td>#B</td></tr><tr><td>#C</td></tr><tr><td>#D</td></tr><tr><td><i>Empty</i></td></tr><tr><td><i>Empty</i></td></tr><tr><td>1kb ladder</td></tr></table><img src=\"https://static.igem.org/mediawiki/2013/6/62/Tn-20130627-GG_R0010pSB1C3_FAILED.jpg\" width=\"450px\" /><br/><img src=\"https://static.igem.org/mediawiki/2013/0/0b/Tn-20130627-GG_R0010pSB1C3_BIS.JPG\" height=\"450px\" /></center></html>}}<html><br/>Finally, Emil prepared the inocula of the product of ligation of R0010 and SAMsynthetase (in pSB1A2: 6 inocula of the 1:1 ligation product and 3 of the 1:3 ligation producta).</html>",
-	"tags" : "EFE"
+	"tags" : "SAMsynthetase-Plac"
 }