Alberta NINT/18 June 2008

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lab work (SD): Results: K102000 + I732018.1 - 134 colonies K102000 + I732018.2 - 125 colonies K102000 (negative control) - 10 colonies Based on sequencing results, 6 colonies from K102000 + I732018.2 plate were inoculated into LB + amp50 culture tubes and incubated at 37 C, shaking, overnight. 10 ul of K102009 overnight tube from 05/06/08 was inoculated into LB + amp50 culture tube and incubated at 37 C, shaking, overnight to create fresh cultures for making a glycerol stock.

For Wayne: Extracted K102011/H+X from gel using QIA quick gel extraction protocol and a min-elute spin column. DNA was eluted in 15ul Buffer EB.

lab work (JD): Miniprep of K102005 from O/N Digested K102005 with BamH1 and Nco1 Ran on 2% agarose gel and cut out desired band Gel purified K102005/B+N Ligated our freshly cut K102005/B+N to both our old TA0 that was heated at 50C for 10 minutes and our newly cut TA0 heated at 50C for 10 minutes Transformed XL1-B cells with these new K102007s Plated K102007 cells Counted number of colonies on K102003 plates from yesterday: 5 on K102003(1) and 2 on K102003(2) and 2 on negative plate Created 7 O/N of these K102003 colonies