法国vilber凝胶&化学发光成像系统 360实验室相关交流-蚂蚁淘在线

法国vilber凝胶&化学发光成像系统 360实验室

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YeastRNAMiniprep

growcellstomid-logphase(10-20ml)(ortakesamplesfromtimecourses,5-25ml)
spincells,discardsupernatant
washcellsoncewith1mlcoldRNAbuffer
freezepelletindryice(andthan@-80°C)
thawcellsonice
add100-120µlicecoldRNA-buffer,resUSPendbyvortexing
add1/2vol.acidwashedglassbeads
vortexathighestlevel3minincoldroomoruseFastPrepmachine(20sec.,level4.5)
add450µlRNA-buffer-SDS(RT),vortexbriefly
add450µlequilibratedPhenol
vortexathighestlevel3minincoldroom
spinfullspeed10minincoldroom
transferupperphasetofreshtube(donottakeanyoftheinterface!)
add300µlequilibratedPhenol,vortex
add300µlChloroform,vortex
spin2min,extractupperphaseonceagainwithChloroform
add20µl4MNaCl,1mlEthanol
precipitate30minat-20to-80°C
spinfullspeed10min
washpelletwith150µl70%Ethanol
airdrypellet
resuspendin30-50µlH₂O,storeat-20to-80°C

Buffers:(Allbuffercomponentsandstock-solutions,excepttheTris-stock-solution,areDEPCtreatedandautoclaved)RNA-buffer:50mMTris(HCl)pH7.4;100mMNaCl;10mMEDTA[for100ml:5ml1MTrispH7.4;2.5ml4MNaCl;2ml0.5MEDTA]RNA-buffer-SDS:RNA-buffer+1.3%SDS

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法国vilber凝胶&amp;化学发光成像系统 360实验室

法国vilber凝胶&化学发光成像系统 360实验室