Team:Calgary/7 July 2009
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*Today I did a colony PCR with p Taq and Biobrick primers (BBK-CP-F and BBK-CP-R) to see if we were able to clone in either the J13002 promoter or the B0015 terminator to our gene of interest, BBK LuxOD47E. PCR products were run on a 1% agarose gel with LuxOD47E BBK as a size control and ddH2O as a negatve control. Gel if pictures below. Lanes 1-4 are J13002-LuxOD47E and Lanes 5-8 are LuxOD47E-B0015. Lane 9 is LuxOD47E BBK, lanes 10 and 11 are left blank and Lane 12 is a negative control with ddH2O. | *Today I did a colony PCR with p Taq and Biobrick primers (BBK-CP-F and BBK-CP-R) to see if we were able to clone in either the J13002 promoter or the B0015 terminator to our gene of interest, BBK LuxOD47E. PCR products were run on a 1% agarose gel with LuxOD47E BBK as a size control and ddH2O as a negatve control. Gel if pictures below. Lanes 1-4 are J13002-LuxOD47E and Lanes 5-8 are LuxOD47E-B0015. Lane 9 is LuxOD47E BBK, lanes 10 and 11 are left blank and Lane 12 is a negative control with ddH2O. | ||
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- | [[Image:2009.07.07J13002-LuxOD47E+LuxOD47E-B0015.jpg]] | + | [[Image:2009.07.07J13002-LuxOD47E+LuxOD47E-B0015.jpg|350px]] |
*From this gel it looks like the cloning of J13002 may have been successful as the band sin the first four lanes appear slighlty higher than the band in the size control lane. We will make overnight cultures of these colonies, isolate plasmid and send the colony with the best concentration for DNA sequencing. | *From this gel it looks like the cloning of J13002 may have been successful as the band sin the first four lanes appear slighlty higher than the band in the size control lane. We will make overnight cultures of these colonies, isolate plasmid and send the colony with the best concentration for DNA sequencing. |
Revision as of 02:12, 14 September 2009
UNIVERSITY OF CALGARY