Team:UNITN-Trento/Notebook/Labposts/07/15

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Latest revision as of 10:35, 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"}

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+{"date" : "2013-07-04","author" : "emil","title" : " Re-Inocula of the plate with the product of ligation and purification of the first plates ","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" : "K832024-K823026-E0840"}
-	"date" : "2013-07-18",
-	"author" : "fabio-bruno",
-	"title" : " blue and red light sensors!!",
-	"content" : "<html>  today we performed the miniprep protocol on our inocula. yealds are: <br>006 a: 169,3 ng/ul; <br>006 b: 204,4ng/ul; <br>016 a: 204,9 ng/ul; <br>016 b: 173,7 ng/ul; <br>030 a: 276ng/ul; <br>030 b: 375,8ng/ul; <br>082 a: 173,1ng/ul; <br>082 b: 225,9 ng/ul; <br>After that we screened 006b, 016a,082 and 030.<br></html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|gel 1|<html><center><img src=\"https://static.igem.org/mediawiki/2013/3/3f/Tn-2013_a.jpg\" width=\"450px\" /></center></html>}}<html></html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|gel 2|<html><center><img src=\"https://static.igem.org/mediawiki/2013/1/18/Tn.2013_B.jpg\" width=\"450px\" /></center></html>}}<html>As we can see from the gels, we didn’t confirm the presence of any of them!! Now we are going to do a second set of inocula!</html>",
-	"tags" : " YF1_FixJ - FixK2- ho1_PcyA_cph8-OmpR"
-}