Alberta NINT/18 August 2008


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lab work (SD):

            Extracted pSB1A3.1 and pSB1A3.3 from the gel from 15/08/08 using QIAquick gel extraction protocol.
            Incubated at 65 C for 8 minutes, vortexed twice for 10 sec. Eluted in 30ul of Buffer EB and let 
            stand for 2 min prior to centrifuging to increase the concentration.
            Sequenced TA2.1, TA2.7, TA3.1, TA3.2, TA6.1 and TA7.2 again using an adapted sequencing protocol.  
            Combined DNA, Vf primer, Sequencing buffer, Pellet Paint and MQ H20, heated at 98 C for 5 min.
            Added Big Dye mix and cooled on ice for 2 min.
            Proceeded with sequencing:
                     [96 C for 10 sec
                      50 C for 20 sec
                      60 C for 4 min] X30
                       4 C hold

lab work (JD):

           Glycerol stock of K102042, K102052, K102062 and K102072
           Transformed XL1-B cells with K102041, K102051, K102061, K102071
           Co-transformed XL1-B cells with K102041+K102042, K102051+K102052, K102061+K102062, K102071+K102072
           Plated transformed cells and incubated overnight