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

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(Created page with "{ "date" : "2013-07-23", "author" : "emil", "title" : "The incompetent Bacillus subtilis saga: The LB transformation and amplification of pSpac +GFP", "content" : "<html>I ha...")
 
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{"date" : "2013-07-02","author" : "emil","title" : "Purification of the digestion products (K823026, K823024, E0840) ligation of the GFP in the 2 backbones and transformation of the ligation products ","content" : "<html>This morning I added 1&micro;l of DPN1 to the GFP ligation and 1&micro;l of SAP(alkaline phosphatase) to the 2 backbones.After 1h 30 min I have stopped the reaction by putting the reaction tubes at 80 C&deg;.Afterwards I have purified the products following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Promega-PCR-Gel\"> purification protocol</a>.Then I quantified the products with the following results:</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Quantification|<html><table>    <tr><th>Sample</th><th>Quantification</th></tr><tr><td>BBa_E0840</td><td> 15.8ng/&micro;l</td></tr><tr><td>BBa_K823026</td><td> 37.6ng/&micro;l</td></tr><tr><td>BBa_K823024</td><td> 36.4ng/&micro;l</td></tr></table></html>}}<html>Then I performed the ligation(1:1,1:3,CTRL) of the GFP with the 2 backbone individually and following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Ligation\"> ligation protocol</a>.Finally I transformed 10&micro;l of the ligation protocol in Neb10&beta; and plate them on ampicillin LB agar following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Competent-cells-transformation\"> transformation protocol</a>.</html>","tags" : "E0840-K823026-K823024"}
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"date" : "2013-07-23",
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"author" : "emil",
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"title" : "The incompetent Bacillus subtilis saga: The LB transformation and amplification of pSpac +GFP",
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"content" : "<html>I have transformed 3 groups of cells with 500 ng of pSpac+pLac+RFP,I grew Bacillus in LB and when it reached OD of 1.1 I added the DNA to 400&micro;l of cells,I let grow for 1 hour and then I plated the cell on Kanamicin plates.Afterwards I transformed Neb10&beta; with 100 ng of K823026+E0840 following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Competent-cells-transformation\">transformation protocol</a>.</html>",
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"tags" : "Bacillus subtilis-pSpac+GFP"
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}
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Latest revision as of 10:33, 3 October 2013

{"date" : "2013-07-02","author" : "emil","title" : "Purification of the digestion products (K823026, K823024, E0840) ligation of the GFP in the 2 backbones and transformation of the ligation products ","content" : "This morning I added 1µl of DPN1 to the GFP ligation and 1µl of SAP(alkaline phosphatase) to the 2 backbones.After 1h 30 min I have stopped the reaction by putting the reaction tubes at 80 C°.Afterwards I have purified the products following the purification protocol.Then I quantified the products with the following results:

Quantification
SampleQuantification
BBa_E0840 15.8ng/µl
BBa_K823026 37.6ng/µl
BBa_K823024 36.4ng/µl
Then I performed the ligation(1:1,1:3,CTRL) of the GFP with the 2 backbone individually and following the ligation protocol.Finally I transformed 10µl of the ligation protocol in Neb10β and plate them on ampicillin LB agar following the transformation protocol.","tags" : "E0840-K823026-K823024"}