Alberta NINT/7 July 2008

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[[Image:gel.jpg]]
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[[Team:Alberta_NINT/Notebook | < Back to notebook]]
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[[Alberta_NINT/4_July_2008 | < Previous entry]] | [[Alberta_NINT/8_July_2008 | Next entry >]]
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lab work (SD):
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            K102022.1 becomes K102022 and K102032.1 becomes K102032.
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            Transformed XL1-B cells with K102022---------------LB + amp50 + IPTG + X-Gal + 20% glucose
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                                          K102032---------------LB + amp50 + IPTG + X-Gal + 20% glucose
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                                          K102032 + K102011-----LB + amp50 + kan35 + IPTG + X-Gal + 20% glucose
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                                          K102032 + K102034-----LB + amp50 + kan35 + IPTG + X-Gal + 20% glucose
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                                          K102032 + K102035-----LB + amp50 + kan35 + IPTG + X-Gal + 20% glucose
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                                          K102032 + K102036-----LB + amp50 + kan35 + IPTG + X-Gal + 20% glucose
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            Negative control plates are arabinose - and test plates are arabinose +
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lab work (JD):
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              From cell counts on plates it is obvious that ligation did not work
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              Suspicious colony on pSB4A5 plate, mini prep of pSB4A5
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              Restriction Digest of pSB4A5 with EcoR1 and Pst1
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              Mini-elute of K102003/E+P and K102006/E+P
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              Ran gel @ 100V and 2% agarose
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              From gel image it may be that we don't really have pSB4A5 but we will proceed as if it
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              is the correct plasmid.
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              Gel purification of pSB4A5/E+P
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              [[Image:annotated6.jpg]]

Latest revision as of 22:57, 11 October 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
              Annotated6.jpg
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