Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/03/11: Difference between revisions
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Rene M Davis (talk | contribs) |
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==Entry title== | ==Entry title== | ||
came in around 10am to check plates. some grew, picked colonies and grew up in lb amp | came in around 10am to check plates. some grew, picked colonies and grew up in lb amp | ||
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miniprepped 8-1, 8-2, 8-3 | |||
colony PCR for ligation of Pconst - B0034 - C0061 | |||
10 colonies | |||
picked, streaked on amp plate, swirled in 15ul water, heated at 95deg for 5 minutes, spun at 12g for 1 minute, added 'sup' to 15ul GoTAQ MM + biobrick primers | |||
controls: 8-1 miniprep and Pconst+RFP miniprep | |||
<br> | |||
Growing the senders! | |||
picked 3 colonies from each BL21 transformation plate and growing up in 2ml LB+amp | |||
grew from 10am to 6 pm | |||
sat on my bench until 7pm | |||
added 1ml of culture to 4ml of media in round bottom tubes | |||
shaking at 37 | |||
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picked some colors without promoters from 2011 plate | |||
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ligations didn't have colonies yet | ligations didn't have colonies yet |
Revision as of 13:52, 28 May 2013
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Entry titlecame in around 10am to check plates. some grew, picked colonies and grew up in lb amp
colony PCR for ligation of Pconst - B0034 - C0061 10 colonies picked, streaked on amp plate, swirled in 15ul water, heated at 95deg for 5 minutes, spun at 12g for 1 minute, added 'sup' to 15ul GoTAQ MM + biobrick primers controls: 8-1 miniprep and Pconst+RFP miniprep
Growing the senders! picked 3 colonies from each BL21 transformation plate and growing up in 2ml LB+amp grew from 10am to 6 pm sat on my bench until 7pm added 1ml of culture to 4ml of media in round bottom tubes shaking at 37
ligations didn't have colonies yet came in a 7PM here's what grew from what was picked this morning:
some of the plates have more colonies. will run colony PCR to see what might be positive if these mini preps don't look good.
2 X & P
picked more colonies from ligations and parts i pulled out yesterday transformed some of the mini preps from today into BL21 2ul of mini prep into 10ul water added 20ul BL21 cells incubated on ice for 8-10 minutes heat shocked at 42deg for 45sec iced for 2 minutes plated K082035 (2-2-2) K084007 (4-2-1 and 4-2-2) K145254 (7-2-1)
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