Team:UNITN-Trento/Notebook/Labposts/07/12

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Revision as of 08:24, 3 October 2013

{ "date" : "2013-07-04", "author" : "emil", "title" : " Re-Inocula of the plate with the product of ligation and purification of the first plates ", "content" : "I purified the inocula done the day before following the purification protocol.Then I quantified the results with the following results:

Quantification

Sample	Quantification
K823026+E0840 1:3 A	409.4ng/µl
K823026+E0840 1:3 B	401.0ng/µl
K823026+E0840 1:1 C	733.4ng/µl
K823026+E0840 1:1 D	489.3ng/µl
K823024+E0840 1:1 E(succesful)	281ng/µl
K823024+E0840 1:3 F	341.1ng/µl
K823024+E0840 1:3 G	220.3ng/µl
K823024+E0840 1:1 error	129.3ng/µl

Then I screened the purified products following the digestion protocol.These are the results of the gel(1%):

Gel order

Sample	Well
Ladder 1kb Fermentas	1
BBa_K823026+BBa_E0840 A	2
BBa_K823026+BBa_E0840 B	3
BBa_K823026+BBa_E0840 C	4
BBa_K823026+BBa_E0840 D	5
BBa_K823024+BBa_E0840 E(the only succesful)	6
BBa_K823024+BBa_E0840 F	7
BBa_K823024+BBa_E0840 G	9

Gel image

As we can see ther are two bands for each well(like expected after the digestion) but only the lower band of the 6 well has the GFP(920 bp) as insert, the other have only the RFP witha promoter.So I decided to repeat the digestion of GFP(an old sample 42.6 ng/µl digested completely) and the same sample of the two backbone exactly like in the previous days.I did the inocula of the new transformation and also of two colonies of the plate that I tooK the E sample from(2 E sample(1:1 024+0840),1 1:1 024+0840,2 1:3 024+0840,2 1:1 026+0840, 2 1:3 026+0840).", "tags" : "K832024-K823026-E0840" }

@@ Line 1: / Line 1: @@
 {
-	"date" : "2013-07-10",
+	"date" : "2013-07-04",
-	"author" : "fabio",
+	"author" : "emil",
-	"title" : " ripenator v1.0 ",
+	"title" : " Re-Inocula of the plate with the product of ligation and purification of the first plates ",
-	"content" : "<html>  this morning, Bruno is not available, so I will take care of the ripenator. I have to replace all the cultures with new LB and new bacteria : I’m gonna put 300 ml of LB in each beutas, then connect immediately a beuta with LB and 3ml of not-induced bacteria to the control banana!! After that, I will dissolve 78 ul of MESA in the second beuta and connect to the blocked banana! As far as the ethylene sample is concerned, I have to put 3 ml of bacteria in the last beuta, wait for the OD lecture to reach the value of 0,5 and then induce the culture with Arabinose to produce Ethylene! Finally I can connect it with the mature banana. </html>",
+	"content" : "<html>I purified the inocula done the day before following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#miniprep\">purification protocol</a>.Then I quantified the results 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>K823026+E0840 1:3 A</td><td>409.4ng/&micro;l</td></tr><tr><td>K823026+E0840 1:3 B</td><td>401.0ng/&micro;l</td></tr><tr><td>K823026+E0840 1:1 C</td><td>733.4ng/&micro;l</td></tr><tr><td>K823026+E0840 1:1 D</td><td>489.3ng/&micro;l</td></tr><tr><td>K823024+E0840 1:1 E(succesful)</td><td>281ng/&micro;l</td></tr><tr><td>K823024+E0840 1:3 F</td><td>341.1ng/&micro;l</td></tr><tr><td>K823024+E0840 1:3 G</td><td>220.3ng/&micro;l</td></tr><tr><td>K823024+E0840 1:1 error</td><td>129.3ng/&micro;l</td></tr></table></center></html>}}<html>Then I screened the purified products following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Digestion\">digestion protocol</a>.These are the results of the gel(1%):</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_K823026+BBa_E0840 A</td><td>2</td></tr><tr><td>BBa_K823026+BBa_E0840 B</td><td>3</td></tr><tr><td>BBa_K823026+BBa_E0840 C</td><td>4</td></tr><tr><td>BBa_K823026+BBa_E0840 D</td><td>5</td></tr><tr><td>BBa_K823024+BBa_E0840 E(the only succesful)</td><td>6</td></tr><tr><td>BBa_K823024+BBa_E0840 F</td><td>7</td></tr><tr><td>BBa_K823024+BBa_E0840 G</td><td>9</td></tr></table></center></html>}}{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Gel image|<html> <img src=\"https://static.igem.org/mediawiki/2013/7/77/Tn-20130704-ET-minscreeningLigation.jpg\" width=\"450\" /></html>}}<html>As we can see ther are two bands for each well(like expected after the digestion) but only the lower band of the 6 well has the GFP(920 bp) as insert, the other have only the RFP witha promoter.So I decided to repeat the digestion of GFP(an old sample 42.6 ng/&micro;l digested completely) and the same sample of the two backbone exactly like in the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Notebook#tn-post-2013-07-01-emil\"> previous days</a>.I did the inocula of the new transformation and also of two colonies of the plate that I tooK the E sample from(2 E sample(1:1 024+0840),1 1:1 024+0840,2 1:3 024+0840,2 1:1 026+0840, 2 1:3 026+0840).</html>",
-	"tags" : " ripenator-mesa-ethylene "
+	"tags" : "K832024-K823026-E0840"
 }