布兰特（1953年出生的德国足球运动员）_
来自 : 蚂蚁淘
Materials:
- TENSsolution:
- 10mMTris(pHto7.5)
- 1mMethylenediaminetetraaceticacid(pHto8.0todissolve)
- 0.1Nsodiumhydroxide
- 0.5%sodiumdodecylsulfate
- 3MSodiumacetate,pH5.2
- Pre-chilled(at-20degreesC)100%ethanol
- 70%Ethanol
- Distilledwater
- Overnightbacterialculture(LAB5)
Supplies:- MicroPipetterandtips
- Vortexmixer
- Microcentrifugeandtubes
Procedures:- Spin1.5mlofovernightbacterialculturefor30-60secondsinamicrocentrifuge.(observation:bacteriaformapelletatthebottomofthetube.)
- Decantsupernatant,leaving50-100ulinthetube.
- VortextoresUSPendthebacteriapelletcompletely.
- Add300ulofTENSsolution.
- Vortexfor5secondstomix.(observation:thecontentsofthetubeshouldbecomeslimy.)
- Add150ulofthesodiumacetate.
- Vortexfor5secondstomix.
- Spinfor2minutesinamicrocentrifuge.(observation:awhitepellet,containingbacterialdebris,isformedatthebottomofthetube.)
- Transfersupernatanttoafreshtube.
- Add0.9mlofpre-chilled100%ethanol.
- Spinfor5minutesinamicrocentrifuge.(observation:awhitepellet,containingplasmidDNAandbacterialRNA,isformedatthebottomofthetube.)
- Discardsupernatantandadd1mlof70%ethanol.
- Discardtheethanolandaddanother1mlof70%ethanol.
- Withdrawanddiscardasmuchliquid(ethanol)aspossIBLethenair-drythepellet.
- Resuspendthepelletin30ulofdistilledwaterandkeepat4degreesCor-20degreesC.
Results:- PlasmidDNAisnowreadyforestimationofDNAconcentration(LAB4)followedbyrestrictiondigest(LAB3).
- Typicalyieldis2-3ug.
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