Team:Michigan/Pili Expression
From 2010.igem.org
(Difference between revisions)
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==9/2/2010== | ==9/2/2010== | ||
+ | ''Kevin'' | ||
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Electroporation of pBAD+FimB into K12 and ΔFimB::kan cells. | Electroporation of pBAD+FimB into K12 and ΔFimB::kan cells. | ||
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==9/7/2010== | ==9/7/2010== | ||
+ | ''Kevin'' | ||
Started overnight cultures of 1:1000 dilutions of K12 and ΔFim::kan, both with the pBAD+FimB plasmid. | Started overnight cultures of 1:1000 dilutions of K12 and ΔFim::kan, both with the pBAD+FimB plasmid. | ||
==9/8/2010== | ==9/8/2010== | ||
+ | ''Kevin'' | ||
Created frozen stocks of K12 and ΔFim::kan w/the plasmid from overnight cultures. Stored them in box 1 in the ERB -20C freezer. | Created frozen stocks of K12 and ΔFim::kan w/the plasmid from overnight cultures. Stored them in box 1 in the ERB -20C freezer. | ||
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==9/9/2010== | ==9/9/2010== | ||
+ | ''Kevin'' | ||
Cryopreserved frozen stocks of K12 and ΔFimB::kan w/the plasmid. | Cryopreserved frozen stocks of K12 and ΔFimB::kan w/the plasmid. | ||
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==9/10/2010== | ==9/10/2010== | ||
+ | ''Kevin'' | ||
Ran gel of previous day's digest, unfortunately the gel was inconclusive. There is an undetermined error with the gel electrophoresis machine. | Ran gel of previous day's digest, unfortunately the gel was inconclusive. There is an undetermined error with the gel electrophoresis machine. | ||
==10/2/2010== | ==10/2/2010== | ||
+ | ''Kevin'' | ||
Prepared tubes with K12 and ΔFim (Both w/insert) for an agglutination assay. We added algae and arabinose at an OD600 of 0.8 and then let the tubes grow | Prepared tubes with K12 and ΔFim (Both w/insert) for an agglutination assay. We added algae and arabinose at an OD600 of 0.8 and then let the tubes grow | ||
==10/3/2010== | ==10/3/2010== | ||
+ | ''Kevin'' | ||
We were able to observe pellets in the tubes containing ΔFimB w/insert. This is exciting, but there are several things to note: | We were able to observe pellets in the tubes containing ΔFimB w/insert. This is exciting, but there are several things to note: | ||
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==10/7/2010== | ==10/7/2010== | ||
+ | ''Kevin'' | ||
Ran flocculation assay. | Ran flocculation assay. | ||
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*In between measurements, the tubes were allowed to sit at room temperature. It may be more beneficial to put the tubes in the incubator shaker, and take measurements less frequently. | *In between measurements, the tubes were allowed to sit at room temperature. It may be more beneficial to put the tubes in the incubator shaker, and take measurements less frequently. | ||
*If we wait 2-3 hrs for the arabinose to fully induce and then start taking measurements, we should get better results. | *If we wait 2-3 hrs for the arabinose to fully induce and then start taking measurements, we should get better results. | ||
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+ | Marc ran another qualitative assay, with positive results. | ||
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+ | [[Image:Floc_tubes10_8.jpg|400px]] | ||
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Revision as of 02:27, 11 October 2010