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Restriction Enzyme Accessibility Protocol in Mammalian Cells

HI Hogune Im

Published: Oct 20, 2011 DOI: 10.21769/BioProtoc.147 Views: 11977

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Abstract

This protocol describes a method to indirectly assess chromatin structure by using restriction enzyme in mammalian cells, modified from Current Protocols.

Materials and Reagents

  1. Tris
  2. NaCl
  3. MgCl2
  4. DTT
  5. EDTA
  6. SDS
  7. Chloroform
  8. PBS
  9. NP-40
  10. 70% EtOH
  11. NaAc
  12. Phenol/chloroform
  13. Digest buffer (NEB)
  14. Restriction enzyme (NEB)
  15. TE (Tris 10 mM, EDTA 1 mM) buffer
  16. Proteinase K (Promega corporation, catalog number: V3021 )
  17. NP-40 (United States biological corporation, catalog number: N3500 )
  18. Lysis buffer (see Recipes)
  19. Wash buffer (see Recipes)
  20. Stop solution (see Recipes)

Equipment

  1. Type B dounce homogenizer
  2. Centrifugation
  3. Falcon 2059 tube

Procedure

Copyright: © 2011 The Authors; exclusive licensee Bio-protocol LLC.

Category

Molecular Biology > DNA > DNA-protein interaction

Cell Biology > Cell structure > Chromosome

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