From 2013.igem.org
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| - | { | + | {"date" : "2013-07-19","author" : "emil-viola","title" : "The incompetent Bacillus subtilis saga OD(io)","content" : "<html>We have tied to grow B.subtilis in the minimal medium (Cambridge protocol and Mansy's friend's protocol) with no evidence of growth, in the previous days we have observed that B. subtilis grow in LB so we have tried to transform B. subtilis in LB following the LMU munich protocol modified.We transformed 2 groups of cells with the construct K823026+GFP but we mistaken the right Kanamicin concentration of the stock solution(10mg/ml)so no the following day ther were no colonies on the plates.</html>","tags" : "Bacillus subtilis"} |
| - | "date" : "2013-07-04",
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| - | "author" : "emil",
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| - | "title" : " Re-Inocula of the plate with the product of ligation and purification of the first plates ",
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| - | "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/µl</td></tr><tr><td>K823026+E0840 1:3 B</td><td>401.0ng/µl</td></tr><tr><td>K823026+E0840 1:1 C</td><td>733.4ng/µl</td></tr><tr><td>K823026+E0840 1:1 D</td><td>489.3ng/µl</td></tr><tr><td>K823024+E0840 1:1 E(succesful)</td><td>281ng/µl</td></tr><tr><td>K823024+E0840 1:3 F</td><td>341.1ng/µl</td></tr><tr><td>K823024+E0840 1:3 G</td><td>220.3ng/µl</td></tr><tr><td>K823024+E0840 1:1 error</td><td>129.3ng/µl</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/µl 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>",
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| - | "tags" : "K832024-K823026-E0840"
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| - | } | + | |
Latest revision as of 10:42, 3 October 2013
{"date" : "2013-07-19","author" : "emil-viola","title" : "The incompetent Bacillus subtilis saga OD(io)","content" : "We have tied to grow B.subtilis in the minimal medium (Cambridge protocol and Mansy's friend's protocol) with no evidence of growth, in the previous days we have observed that B. subtilis grow in LB so we have tried to transform B. subtilis in LB following the LMU munich protocol modified.We transformed 2 groups of cells with the construct K823026+GFP but we mistaken the right Kanamicin concentration of the stock solution(10mg/ml)so no the following day ther were no colonies on the plates.","tags" : "Bacillus subtilis"}
"
