IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/06/16

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12:30 PM

Promoter BBa_J23119: had growth on both plates for new and used cuvettes. Took 1 colony from new cuvette plate (50 ul plated) and inoculated 5 mL LB broth.

Gas Vesicles BBa_I750016:

made glycerol stock: .9 mL 10% glycerol and .9 mL culture in tube in -80°C freezer.

Miniprepped (promega kit) remaining sample and nanodropped (432.57 ng/ul)


used 100ng DNA as template

Ingredient amount/reaction mastermix (3 reactions)
10X buffer 2 ul 6 ul
10 mM dNTPs .5 ul 1.5 ul
DNA .5 ul added individually to each reaction
F Primer .2 ul .6 ul
R Primer .2 ul .6 ul
Polymerase .4 ul 1.2 ul
H2O 16.2 ul 48.6 ul

19.5 ul mastermix and .5 ul template DNA to each reaction (except negative control which gets .5 ul H2O instead).

Samples: BBa_I750016, negative control.

Program: BBa6Kb

Size: ~6Kb

Primers: Biobrick sequencing forward and reverse.