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文献和实验Preparation of nuclear extract and cytoplasmic extract
an aliquot with Trypan Blue and check for >90% lysis on microsope.5.Spin at 2000 rpm for 15 minutes in J-6 to pellet nuclei.6.Save the supernatant for S100 extract (see below).7.Transfer the nuclear pellet to a glass beaker containing a stir bar and add
Largescale nuclear extract preparation
,Brenner D (1990)A simplified method for the preparation of transcrip-tionally active liver nuclear extracts.DNA Cell Biol.Vol.10,P.777-781 I.Homogenize cells or tissue 1.Wash commercially available HeLa cells (30g)or minced in 3mm pieces
Preparation of HeLa Nuclear Extracts
Promoter-specific in vitro transcription by RNA polymerase II is now achievable with cell extracts prepared from a variety of animal and plant sources (1 –3 ). For extracts prepared from mammalian cells, HeLa cells provide many advantages
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