Nucleosomes by Paul M. Wassarman and Roger D. Kornberg (Eds.)

By Paul M. Wassarman and Roger D. Kornberg (Eds.)

The significantly acclaimed laboratory common, Methods in Enzymology, is among the so much hugely revered courses within the box of biochemistry. in view that 1955, each one quantity has been eagerly awaited, usually consulted, and praised via researchers and reviewers alike. The sequence comprises a lot fabric nonetheless proper at the present time - actually a necessary booklet for researchers in all fields of lifestyles sciences

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Supernatants are transferred to fresh tubes and kept on ice. It is convenient to perform the subsequent steps on 1-ml aliquots in Eppendorf microcentrifuge tubes. Preparation of Antibody- Immunobead Complex A 1-ml aliquot of Immunobeads is washed twice with PBS and resuspended in 1 ml of PBS; a 10-see centrifugation in a microfuge at room temperature is sufficient to pellet the beads in this and all steps following. 5 mg/ml per 100/~1 of bead suspension). The mixture is incubated on ice for 2 hr with occasional mixing.

Coli strain BMH 72-19-18 and purified exactly as described by Fowler and Zabin. 9 The purified protein is stored in aliquots at -80*. 6 A. D. Riggs, H. Suzuki, and S. Bourgeois, J. Mol. Biol. 48, 67 (1970). 7 D. Lcvens and P. M. Howley, MoL Cell. Biol. S, 2307 (1985). 8 B. Muller-Hill and J. Kania, Nature (London) 249, 561 (1974). 9 A. V. Fowler and I. Zabin, J. Biol. Chem. 258, 14354 (1983). 36 CHROMATIN AND NUCLEOSOMES: PREPARATIVE TECHNIQUES [3] Rabbit anti-fl-galactosidase antibodies (Cappel Laboratories, Malvern PA) are repurified by binding to E.

L. DePamphilis, J. Virol. 48, 281 (1983). 29 L. C. Tack, J. H. Whright, and E. G. Gurney, J. Virol. 58, 635 (1986). 3o j. L. Workman and J. P. Langmore, Biochemistry 24, 7486 (1985). 26 CHROMATIN AND NUCLEOSOMES: PREPARATIVE TECHNIQUES [3] [3] I s o l a t i o n o f Y e a s t P l a s m i d C h r o m a t i n By ANN DEAN, DAVID S. PEDERSON,and ROnERT T. SIMPSON Introduction Recombinant DNA technology has allowed insight into the mechanisms of control of gene expression in eukaryotic cells (see Ref.

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