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Contents 1 Lac Operon Control 2 Vary RBS 3 Chitin 3.1 Chitin Synthesis 3.2 Chitin Staining (Calcofluor) 4 Apoptosis 4.1 Live/Dead Staining aka Viability/Cytotoxicity Assay 5 Keio Collection - KnockOuts 5.1 ChiA(Chitinase) Knockout (may not be necessary) 5.2 ydgg/tqsA (AI2-Transporter) Knockout

Lac Operon Control

http://openwetware.org/wiki/IPTG

Vary RBS

http://partsregistry.org/Ribosome_Binding_Sites/Prokaryotic/Constitutive/Community_Collection

Chitin

Chitin Synthesis

chitin synthase gene (chs3): http://www.ncbi.nlm.nih.gov/gene/852311

Chitin Staining (Calcofluor)

Does not stain E Coli - http://www.sigmaaldrich.com/etc/medialib/docs/Fluka/Datasheet/18909dat.pdf

Stains Biofilm Polysaccharides? - http://www.sciencedirect.com/science?_ob=MImg&_imagekey=B6V73-3YKKH2J-4S-1&_cdi=5831&_user=965532&_pii=0043135494003399&_orig=search&_coverDate=08%2F31%2F1995&_sk=999709991&view=c&wchp=dGLbVzz-zSkzS&md5=fedcb287ed9c9dafca2dbb4f2bc053a4&ie=/sdarticle.pdf

Inhibits Synthase - http://www.ncbi.nlm.nih.gov/pmc/articles/instance/40653/

Apoptosis

registry: http://partsregistry.org/Cell_death

Live/Dead Staining aka Viability/Cytotoxicity Assay

DMAO(Live), EtD-III(Dead) - http://www.promokine.info/products/cell-analysis/cell-staining-reagents-related-products/, http://www.biotium.com/product/product_info/Protocol/30027.pdf

Keio Collection - KnockOuts

http://www.shigen.nig.ac.jp/ecoli/strain/top/top.jsp

ChiA(Chitinase) Knockout (may not be necessary)

summary of e coli knockouts: http://openwetware.org/wiki/Escherichia_coli/Knockouts

chitinase gene (chiA): http://www.ncbi.nlm.nih.gov/gene/947837

chiA-knockout strain e coli: http://ecoli.naist.jp/GB6/info.jsp?id=JW3300

ydgg/tqsA (AI2-Transporter) Knockout

KO -> ~7000x biofilm thickness, ~500x biomass - http://jb.asm.org/cgi/content/short/188/2/587