Minnesota/25 July 2008

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Revision as of 15:23, 25 July 2008 by Emartin9808 (Talk | contribs)
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1. No growth on plates: Experiments on 07-24-08 failed to show growth on Kanamycin resistant plates.
2. Redoing ligation: Using left over double digest from 07-24-08, But increase the amount of ligase in the reaction and performing the ligations @ 60C which is the optimal temperature for the ligase.
Parts 10x Buffer H20 Base Vector Insert DNA T4 DNA Ligase Total Volume
BV + Tet1 4.0uL 11.5uL 3.5uL 17.0uL 4.0uL 40.0uL
BV + Lac2 4.0uL 13.5uL 3.5uL 15.0uL 4.0uL 40.0uL
BV + Lac/LAMBDA 4.0uL 15.5uL 3.5uL 13.0uL 4.0uL 40.0uL
BV + Tet/p22 4.0uL 11.5uL 3.5uL 17.5uL 4.0uL 40.0uL
3. Redoing transformations: Redo with new ligated products by adding much less DNA/plasmid to the TOP10 Cells, b/c if put too much ligated DNA then the cells do not transform properly (as stated by iGEM troubleshooting).
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