Alberta NINT/18 June 2008

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lab work (SD):
lab work (SD):
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               Ran on 2% agarose gel and cut out desired band
               Ran on 2% agarose gel and cut out desired band
               Gel purified K102005/B+N
               Gel purified K102005/B+N
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               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
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               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
               Transformed XL1-B cells with these new K102007s
               Plated K102007 cells
               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
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               Counted number of colonies on K102003 plates from yesterday: 5 on K102003(1) and 2 on K102003(2)  
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              and 2 on negative plate
               Created 7 O/N of these K102003 colonies
               Created 7 O/N of these K102003 colonies
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              [[Image:annotated3.jpg]]

Latest revision as of 22:51, 11 October 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
              Annotated3.jpg