Week of 9/16/12
Performed gel extraction of ligated backbones (both E+P and E+S); failed to show up in
adequate amounts when nanodropped.
Ligated Plasmid+RBS (E+S), SynCG (X+P), and RFP (X+P) at 3:1 and 1:1 ratios of both
2X ligase buffer and promega DNA ligase and performed PCR.
Repeated ligation and performed PCR purification: P+RBS+RFP and P+RBS+SynCG
demonstrate proper amounts of DNA but at low concentrations, suggesting that perhaps
the DNA is not the intended amplicon.
Discovered that roughly half of the previous pregnancy tests now appear positive,
brighter than evaporation lines would be. Some tests for 5 mM DTT (denaturing
solution), 0.5 mM DTT, 0.05 mM DTT, and 0 mM DTT are now positive with the
majority in the latter two groups.
Nanodropped the P+RBS+RFP, P+RBS+SynCG, backbone (E+S), backbone (E+P),
and SynCG all (E+P): only the backbone (E+S) and SynCG all (E+P) showing proper
concentrations. The latter includes the IDT plasmid backbone.
Plated the bbp9 biobricks (both 3:1 and 1:1 ratios), the hCG biobrick (in 3:1, 1:1, and
1:3 ratios), backbone, original BBa_K331022, and a DNA lacking group, producing
thousands of cells on each plate (except for the last), demonstrating that many of the cells
have empty vectors.