Alberta NINT/18 June 2008

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(Difference between revisions)
(New page: lab work (SD): Results: K102000 + I732018.1 - 134 colonies K102000 + I732018.2 - 125 colonies K102000 (negative control) - 10 colon...)
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             For Wayne: Extracted K102011/H+X from gel using QIA quick gel extraction protocol and a min-elute  
             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.
             spin column.  DNA was eluted in 15ul Buffer EB.
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 +
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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
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              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

Revision as of 19:49, 19 June 2008

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
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