CD ChIP-Seq Spike-In Normalization Control

Inquiry

CAT	Specs	Inquiry	Basket
EP007-01	Custom
EP007-02	50 µg

CD ChIP-Seq Spike-In Normalization Control uses Drosophila melanogaster-derived reference chromatin together with a corresponding species-specific spike-in antibody. Adding the same reference control to each mammalian ChIP reaction provides an external basis for measuring technical variation introduced during immunoprecipitation, DNA recovery, amplification, and sequencing. The reference configuration includes two ordering components: EP007-01 Reference Chromatin in a custom format and EP007-02 Species-specific Spike-in Antibody, 50 µg at 1 µg/µl. Reference reaction conditions use 25 µg experimental chromatin, 4 µg antibody of interest, and 2 µg spike-in antibody. Optimize the amount of reference chromatin for the experimental workflow; spike-in reads should account for no more than 5% of the sequencing run. Add reference chromatin and spike-in antibody immediately before immunoprecipitation, perform ChIP with the experimental antibody, map sequencing reads to the experimental and reference genomes, and calculate a normalization factor from reference read counts. For research use only. Not for use in diagnostic procedures.

Storage:

Store each component according to its product label. Avoid repeated freeze/thaw cycles of the antibody.

Specifications:

Features	EP007-01 provides Drosophila melanogaster-derived reference chromatin in a custom format. EP007-02 reference format: 50 µg species-specific spike-in antibody at 1 µg/µl. Uses non-mammalian reference chromatin and a corresponding species-specific antibody. Supports normalization of technical variation across sample processing, immunoprecipitation, DNA recovery, amplification, and sequencing. Compatible with ChIP-qPCR and ChIP-seq workflows.
Application	It is recommended to use this control for: Normalization of ChIP-qPCR and ChIP-seq data Comparison of broad changes in chromatin-associated signals Assessment of technical variation between samples and batches ChIP workflow development and troubleshooting
Species Category	Mammalian experimental samples with Drosophila melanogaster reference chromatin
Sample type	Cross-linked mammalian chromatin prepared for ChIP
Sample amount	Reference reaction conditions: 25 µg experimental chromatin, 4 µg antibody of interest, and 2 µg spike-in antibody. Optimize the amount of reference chromatin for the experimental workflow; spike-in reads should account for no more than 5% of the sequencing run.
Sequencing Platform	Illumina; qPCR