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Restriction Enzyme Accessibility Protocol in Mammalian Cells
Published: Oct 20, 2011 DOI: 10.21769/BioProtoc.147 Views: 12005
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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
- Tris
- NaCl
- MgCl2
- DTT
- EDTA
- SDS
- Chloroform
- PBS
- NP-40
- 70% EtOH
- NaAc
- Phenol/chloroform
- Digest buffer (NEB)
- Restriction enzyme (NEB)
- TE (Tris 10 mM, EDTA 1 mM) buffer
- Proteinase K (Promega corporation, catalog number: V3021 )
- NP-40 (United States biological corporation, catalog number: N3500 )
- Lysis buffer (see Recipes)
- Wash buffer (see Recipes)
- Stop solution (see Recipes)
Equipment
- Type B dounce homogenizer
- Centrifugation
- 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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