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Imprint® Ultra Chromatin Optimization Kit

Kit designed to optimize sonication parameters for ChIP experiments

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NACRES:
NA.54

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General description

The Imprint Ultra Chromatin Optimization Kit provides reagents needed to optimize chromatin sonication/shearing conditions before performing ChIP. It is imperative that chromatin is completely sheared into fragments with a peak around 500 base pairs (bp) for ChIP-qPCR assays, and < 500 bp for ChIP-Seq and ChIP-chip, for maximum specificity and resolution, and to ensure low background. For higher resolution applications like genome-wide location analysis by ChIP-chip or ChIP-Seq, sonication of input chromatin to below 500 bp, with a peak around 250 bp, is critical.

Components

Sufficient reagents are provided to perform three sets of chromatin sonication optimization experiments. Each set would include five different conditions for sonication to optimize the parameters. Reagents are provided for a total of 15 reactions, including preparation of cell lysates from 107 cells/reaction, nuclei lysis, sonication, cross-link reversal and DNA purification.

Legal Information

Imprint is a registered trademark of Merck KGaA, Darmstadt, Germany

Kit Components Also Available Separately

Product No.
Description
SDS
  • I8896IGEPAL® CA-630, for molecular biologySDS
  • S5150Sodium chloride solution, 5 M in H2O, BioReagent, for molecular biology, suitable for cell cultureSDS
  • W4502Water, Nuclease-Free Water, for Molecular BiologySDS
  • A84564-(2-Aminoethyl)benzenesulfonyl fluoride hydrochloride, ≥97.0% (HPLC)SDS
  • P8340Protease Inhibitor Cocktail, for use with mammalian cell and tissue extracts, DMSO solutionSDS
  • R4642Ribonuclease A from bovine pancreas, (Solution of 50% glycerol, 10mM Tris-HCL pH 8.0)SDS

Signal Word

Danger

Hazard Classifications

Acute Tox. 4 Oral - Aquatic Acute 1 - Aquatic Chronic 1 - Eye Dam. 1 - Flam. Liq. 3 - Met. Corr. 1 - Ox. Liq. 1 - Resp. Sens. 1 - Skin Corr. 1B - STOT SE 3

Target Organs

Central nervous system

Storage Class Code

5.1A - Strongly oxidizing hazardous materials

WGK

WGK 3

Flash Point(F)

100.4 °F

Flash Point(C)

38 °C

Regulatory Information

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Henriette O'Geen et al.
BioTechniques, 41(5), 577-580 (2006-12-05)
A single chromatin immunoprecipitation (ChIP) sample does not provide enough DNA for hybridization to a genomic tiling array. A commonly used technique for amplifying the DNA obtained from ChIP assays is ligation-mediated PCR (LM-PCR). However; using this amplification method, we
Genetic Framework for GATA Factor Function in Vascular Biology
Linnemann A.K., et al.
Proceedings of the National Academy of Sciences of the USA, 1, 13641-13646 (2011)
M H Kuo et al.
Methods (San Diego, Calif.), 19(3), 425-433 (1999-12-02)
Chromatin structure plays important roles in regulating many DNA-templated processes, such as transcription, replication, and recombination. Considerable progress has recently been made in the identification of large, multisubunit complexes dedicated to these nuclear processes, all of which occur on nucleosomal
Yoon-A Kang et al.
Molecular and cellular biology, 32(1), 226-239 (2011-10-26)
Developmental and homeostatic remodeling of cellular organelles is mediated by a complex process termed autophagy. The cohort of proteins that constitute the autophagy machinery functions in a multistep biochemical pathway. Though components of the autophagy machinery are broadly expressed, autophagy

Protocols

ChIP-qPCR Data Analysis

Chromatin Immunoprecipitation qPCR for studying gene regulation across conditions.

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