Team:Groningen/Protocols

(Difference between revisions)

Latest revision as of 23:02, 27 October 2010

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-[[CaCl2 Competent Cells]]
+== Protocols ==
-'''Day 1:'''
+[[Team:Groningen/Transformation to E. coli | Transformation to ''E. coli'']]
--Inoculate 5 ml LB( TY)  medium with ''E.coli'' from glycerol stock
+[[Team:Groningen/Normal SDS-PAGE using Tris-Glycine Gels and Electrophoresis Buffer | Normal SDS-PAGE using Tris-Glycine Gels and Electrophoresis Buffer]]
--Grow overnight at 37 oC
+[[Team:Groningen/Protocols for Bacillus subtilis 168 | Protocols for Bacillus subtilis 168]]
-'''Day 2:'''
+[[Team:Groningen/Chaplins | Chaplins]]
--Inoculate 20 ml LB medium with 200 μl overnight culture
+[[Team:Groningen/Protocols for Streptomyces | Protocols for Streptomyces]]
--Grow at 37 oC until OD600 = 0,3-0,5(+/- 2 hours)
+[[Team:Groningen/Protocols for Lactococcus | Protocols for Lactococcus]]
--Spin down 5 minute at 4000 rpm at 4 oC
+[[Team:Groningen/Antibiotics and Concentrationas | Antibiotics and Concentrations ]]
--Resuspend in 10 ml chilled 0,1 M caCl2 (from here, keep on ice! )
+[[PelletPrepGR| Chaplin isolation: cell pellet]]
--Inoculate on ice for 20 minutes
+[[SupernatantPrepGR| Chaplin isolation: supernatant]]
--Spin down as before
+[[ExtractionCellWallsGR| Chaplin isolation: cell wall isolation]]
--Remove supernatant
+[[LysozymePrepGR| Chaplin isolation: lysozyme preperation]]
--Resuspend in 2 ml 0,1 M CaCl2 /10% glycerol
--Divide 100 μl aliquots
--Store competent cells in - 80 oC