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

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(Difference between revisions)

Latest revision as of 10:34, 3 October 2013

{"date":"2013-07-03","author":"viola","title":"cloning of the terminators with EFE","content":" Yesterday's ligation seemed to work and and this evening i'll make some inocula... ","tags":"EFE"}

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+{"date":"2013-07-03","author":"viola","title":"<html>cloning of the terminators with EFE</html>","content":"<html> <a href=\"https://2013.igem.org/wiki/index.php?title=Team:UNITN-Trento/Notebook#tn-post-2013-07-02-viola\">Yesterday's ligation</a> seemed to work and and this evening i'll make some inocula... </html>","tags":"EFE"}
-	"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"
-}