From 2013.igem.org
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| - | { | + | {"date" : "2013-07-04","author" : "fabio-viola","title" : "BACILLUS SUBTILIS COMES BACK TO LIFE!","content" : "<html> today we made glycerol stock tubes from the remaining cultures and from the inocula made yesterday, but before that we collected some bacteria to make other two liquid cultures just to have more glycerol stock tubes to put at -80°</html>","tags" : " Bacillus subtilis "} |
| - | "date" : "2013-07-05",
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| - | "author" : "emil",
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| - | "title" : " Purification of the second run of Inocula of ligation and re-ligation of the digested products ",
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| - | "content" : "<html> I added 1µl of DPN1 to the insert and 1µl of SAP to the backbones.Then I have purified the inocula following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#miniprep\"> miniprep protocol</a>.Afterwards I quantified them toghether with the results of the purification of the digestion products(performed following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Promega-PCR-Gel\"> purification protocol </a>) 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>1:1 024+0840 E</td><td>482.6ng/µl</td></tr><tr><td>1:1 024+0840 E</td><td>700ng/µl</td></tr><tr><td>1:1 024+0840</td><td>373.1ng/µl</td></tr><tr><td>1:3 024+0840</td><td>423.5ng/µl</td></tr><tr><td>1:3 024+0840</td><td>327.8ng/µl</td></tr><tr><td>1:1 026+0840</td><td>293.1ng/µl</td></tr><tr><td>1:1 026+0840</td><td>1243ng/µl</td></tr><tr><td>1:3 026+0840</td><td>766.4ng/µl</td></tr><tr><td>1:3 026+0840</td><td>369.1ng/µl</td></tr><tr><td>GFP</td><td>15.9ng/µl</td></tr><tr><td>K823026</td><td>36.3ng/µl</td></tr><tr><td>K823026</td><td>43.7ng/µl</td></tr></table></center></html>}}<html>Then I digested 800 ng of each sample with EcoR1 HF and Pst1 following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Digestion\">digestion protocol</a> with the following results:</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_K823024+BBa_E0840 A</td><td>3</td></tr><tr><td>BBa_K823024+BBa_E0840 B</td><td>5</td></tr><tr><td>BBa_K823024+BBa_E0840 C</td><td>6</td></tr><tr><td>BBa_K823024+BBa_E0840 D</td><td>7</td></tr><tr><td>BBa_K823024+BBa_E0840 E</td><td>8</td></tr><tr><td>BBa_K823026+BBa_E0840 F</td><td>9</td></tr><tr><td>BBa_K823026+BBa_E0840 G</td><td>10</td></tr><tr><td>BBa_K823026+BBa_E0840 H</td><td>11</td></tr><tr><td>BBa_K823026+BBa_E0840 I</td><td>12</td></tr></table></center></html>}}{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Gel image|<html><img src=\"https://static.igem.org/mediawiki/2013/4/4b/Tn-20130705-ET-Ligation_screening2.jpg\" width=\"450\" /></html>}}<html>As we can see no one of the wells shows the right insert in the lower band(there is always the previous insert the RFP + Lac promoter ~ 1200 bp).At the same time I did the ligation of K823024+E0840 and K823026+E0840 previously digested following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Ligation\"> ligation protocol </a>.<br>N.B. Remember always to spin the ligation buffer before using<br> After that I plated the product of the ligation as done before(1:1,1:3,ctrl for each sample) on Ampicillin added LB.</html>",
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| - | "tags" : "K832024-K823026-E0840"
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| - | } | + | |
Latest revision as of 10:35, 3 October 2013
{"date" : "2013-07-04","author" : "fabio-viola","title" : "BACILLUS SUBTILIS COMES BACK TO LIFE!","content" : " today we made glycerol stock tubes from the remaining cultures and from the inocula made yesterday, but before that we collected some bacteria to make other two liquid cultures just to have more glycerol stock tubes to put at -80°","tags" : " Bacillus subtilis "}
"
