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

From OpenWetWare
Jump to: navigation, search
UIUC ArsR System.jpg UIUC - Arsenic Collection System! Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

6/16/10

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)

PCR

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.