Alberta NINT/7 July 2008

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lab work (SD): K102022.1 becomes K102022 and K102032.1 becomes K102032. Transformed XL1-B cells with K102022---LB + amp50 + IPTG + X-Gal + 20% glucose K102032---LB + amp50 + IPTG + X-Gal + 20% glucose K102032 + K102011-LB + amp50 + kan35 + IPTG + X-Gal + 20% glucose K102032 + K102034-LB + amp50 + kan35 + IPTG + X-Gal + 20% glucose K102032 + K102035-LB + amp50 + kan35 + IPTG + X-Gal + 20% glucose K102032 + K102036-LB + amp50 + kan35 + IPTG + X-Gal + 20% glucose Negative control plates are arabinose - and test plates are arabinose +

lab work (JD): From cell counts on plates it is obvious that ligation did not work Suspicious colony on pSB4A5 plate, mini prep of pSB4A5 Restriction Digest of pSB4A5 with EcoR1 and Pst1 Mini-elute of K102003/E+P and K102006/E+P Ran gel @ 100V and 2% agarose From gel image it may be that we don't really have pSB4A5 but we will proceed as if it               is the correct plasmid. Gel purification of pSB4A5/E+P