实验三 细菌的形态学鉴定
来自 : 蚂蚁淘
LyophilizingCells- Inoculate200mlL-brothsupplementedwithappropriateantibioticswiththebacteriatobelyophilized.
- Incubatethecultureat37°Cwithvigorousshaking(200-250rpm)overnight.
- Add2ml(1/100volume)offormaldehydesolutiontoculture.Incubateat37°Cforonehouronarotaryplatformtokillthecells.
- Aliquot100mloftheculturetoeachoftwo250-mlcentrifugebottles.Recoverthecellsbycentrifugationat7,000rpm,4°C,for15minutes.
- WashthecellsbyresUSPendingeachpelletin100ml(1volume)sterile1xPBS.Pelletthecellsbycentrifugationasbefore.
- Resuspendeachpelletin10ml(1/10volume)sterile1xPBS,andrecombineintoa50-mlconicalbottom,polypropylenecentrifugetube.
- Freezethecontentsofthetubeat-70°Cforapproximatelyonehour,oruntilfrozensolid.Freezethetubeinaninclinedpositiontomaximizesurfaceareaandfacilitatelyophilization.
- Placethefrozentube,withthecaploosened,intheglasslyophilizationvessel.Attachthevesseltothelyophilizer,anddesiccateundervacuumuntildry(maytakeseveraldays).
- Storethetightly-cappedtubeat4°Cinazip-lockplasticbagcontainingdessicant.
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