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From 2008.igem.org

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Revision as of 23:15, 7 August 2008

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Contents 1 Agenda 2 Minutes 3 Action Items 4 Coming Up

Agenda

1. Just some advice from our professors.

Minutes

Present: Dr. Presting, Dr. Gautz, Dr. Callahan, Norman, Krystle, Grace, & Margaret

1. Concerning the Omega Interposon: We have had problems amplifying, so Dr. C suggested that we make it into a BioBrick by simply digesting a BB plasmid with one of the sites also in the polylinker of the Omega Interposon and neatly shoving it in that vector. Preliminary knowledge of what is available makes me want to put O.I. into the ccdB gene. I need to find out if this gene will be disrupted. If this doesn't work, Norman suggested that we synthesize an insert to put it in. For example BglII could be the restriction site engineered into this insert, it is compatible with BamHI, so when the O.I. is inserted, the site will be destroyed.
2. Concerning Ligation:

• We need to use at least 100ng-1ug of insert
• Ethanol Precipitation, from Norman's experience, yields 10 fold less DNA than you start with.
• You must clean up the PCR reaction before digest, the taq will keep adding nucleotides if you do not.
• We should start using better controls for the transformation of our ligation products. As a negative control, use the vector ligated back to itself.

1. Concerning Blue/White Selection

Action Items

• <Person A>: Task

Coming Up

[http://manoa.hawaii.edu/ ][http://manoa.hawaii.edu/ovcrge/ ][http://www.ctahr.hawaii.edu ]

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