Alberta NINT/16 June 2008

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lab work (SD): Isolated DNA (I732018) from 15/06/08 LB + amp culture tubes using QIAprep spin miniprep protocol, washing with Buffer PB to eliminate any potential high nuclease activity and prevent loss of DNA. Set up I732018 for sequencing. Digested I732018 with XbaI + PstI. Ran on 2% agarose gel (110V 50 minutes) and excised the 250 bp fragments from I732018.1, .2 and .4 Extracted DNA from gel usig QIAquick gel extraction protocol. Combined .1 and .4 to give higher concentration of DNA. Ligated K102000 with I732018.1 and with I732018.2 (Negative control K102000).

lab work (JD): Only 2 colonies were found on K102007 positive and negative plates so ligation didn't work Still carried out mini-prep on K102007.1 and K102007.2 from these colonies Sequenced K102007.1 and K102007.2 Since 1/100 dilution of TA0 didn't work well we want to recarry out ligations with 1/10, 1/100 and 1/1000 dilutions Performed the appropriate dilutions Carried out ligations of K102005/B+N and 1/10, 1/100, 1/1000 dilutions of TA0/B+N Transformed XL1-B cells