Team:UNITN-Trento/Notebook/Labposts/07/20

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{"date" : "2013-07-08","author" : "emil","title" : " Purification of Inocula (03/7)","content" : "<html>I verifyed the status of the last inocula(05/7) that resulted a bit strange.</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Inocula|<html><img src=\"\" width=\"450px\" /></html>}}<html>Unfortunately The day before were inoculate uncorrect plates(03/7) so I have purified them following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#miniprep\"> purification protocol </a> these are the results of the quantification.</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Quantification|<html><center><table class=\"tn-sp-table\"><tr><th>Sample</th><th>Quantities</th></tr><tr><td>BBa_K823026+BBa_E0840(1:1) 1</td><td>438.8 ng/&micro;l</td></tr><tr><td>BBa_K823026+BBa_E0840(1:1) 2</td><td>400.7 ng/&micro;l</td></tr><tr><td>BBa_K823024+BBa_E0840(1:1) 3</td><td>343.6 ng/&micro;l(the only succesful)</td></tr></table></center></html>}}<html>Afterwards I screened 800 ng of  the samples following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Digestion\">screening protocol</a> with EcoR1 HF and Pst1, these are the results of the gel:</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(1:1) a</td><td>3</td></tr><tr><td>BBa_K823026+BBa_E0840(1:1) b</td><td>4</td></tr><tr><td>BBa_K823024+BBa_E0840(1:1) </td><td>5</td></tr></table></center></html>}}{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Gel image|<html><img src=\"\" width=\"450px\" /></html>}}<html>As we can see only the third(024) sample shows the insert at the right size of 1000 bp(the lower band), then I did the inocula of the plates of 5/07(1:1,1:2,1:3) and of the old plate that gives right results to amplify 024(1:1).</html>","tags" : "K823024-K823026-E0840"}
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"date" : "2013-07-23",
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"author" : "fabio",
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"title" : " the blue ligation: the never ending story !",
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"content" : "<html> this morning I added Sap to the 016 digestion and dPnl to the other two, then I purified them and quantified'em: <br>O16: 29,8 ng/ul <br>006: 16,2 ng/ul <br>R0010: 6,1 ng/ul <br>Then I started the ligation following the ligation protocol: at the end of it, I transformed 10ul of the ligation in 200 Neb10b and plated.Meanwhile Bruno miniprepped k952003 inocula and the screening was successful!!</html>",
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"tags" : " YF1_FixJ - FixK2- ho1_PcyA_cph8-OmpR- YF1_FixJ_PfixK2_amilGFP "
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Latest revision as of 10:37, 3 October 2013

{"date" : "2013-07-08","author" : "emil","title" : " Purification of Inocula (03/7)","content" : "I verifyed the status of the last inocula(05/7) that resulted a bit strange.

Unfortunately The day before were inoculate uncorrect plates(03/7) so I have purified them following the purification protocol these are the results of the quantification.
Quantification
SampleQuantities
BBa_K823026+BBa_E0840(1:1) 1438.8 ng/µl
BBa_K823026+BBa_E0840(1:1) 2400.7 ng/µl
BBa_K823024+BBa_E0840(1:1) 3343.6 ng/µl(the only succesful)
Afterwards I screened 800 ng of the samples following the screening protocol with EcoR1 HF and Pst1, these are the results of the gel:
Gel order
SampleWell
Ladder 1kb Fermentas1
BBa_K823026+BBa_E0840(1:1) a3
BBa_K823026+BBa_E0840(1:1) b4
BBa_K823024+BBa_E0840(1:1) 5
As we can see only the third(024) sample shows the insert at the right size of 1000 bp(the lower band), then I did the inocula of the plates of 5/07(1:1,1:2,1:3) and of the old plate that gives right results to amplify 024(1:1).","tags" : "K823024-K823026-E0840"}