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Alberta NINT/7 July 2008
From 2008.igem.org
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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
