Description:

KAPA hgDNA Quantification and QC Kit

Quality matters.

KAPA hgDNA Quantification and QC Kits contain all the reagents needed for the qPCR-based quantification and quality assessment of human genomic DNA samples prior to NGS library construction.

Kits contain KAPA SYBR^® FAST qPCR Master Mix, optimized for high-performance SYBR Green I-based qPCR, as well as a pre-diluted set of DNA standards and primer premixes targeting different portions of a highly conserved single-copy human locus. Absolute quantification is achieved with the primer pair defining the shortest fragment, whereas the additional primers are used to derive information about the amount of amplifiable template in the DNA sample. Quality scores (or Q-ratios) generated with the kit may be used to predict the outcome of library construction, or tailor workflows for samples of variable quality, particularly FFPE DNA.

The method is easy to automate and can be applied to any process or workflow that requires accurate quantification of dilute DNA samples, or samples that may contain a high proportion of DNA that is recalcitrant to PCR amplification.

Download our KAPA hgDNA Quantification and QC Data Analysis Template.

For Research Use Only. Not for use in diagnostic procedures.

Product Highlights

Obtain concentration and quality information with a single assay

• Allows for accurate quantification of dilute DNA samples
• Quantification with an additional primer pair provides a Q-ratio that is indicative of sample quality

Employ quality scores in the analysis of NGS library construction workflows

• Q-ratios can provide valuable insights into the bottlenecks in NGS library construction workflows
• For FFPE samples, library and sequence quality is primarily limited by inefficient conversion of input DNA to adapter-ligated library

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0.4 yielded libraries that met minimum sequence quality requirements. Data on file.\">0.4 yielded libraries that met minimum sequence quality requirements. Data on file.\">

Obtain actionable data for sample preparation

• Q-ratios correlate with key sequencing metrics such as duplication rates and mean target coverage
• FFPE samples with a Q score >0.4 yield libraries of acceptable quality when processed in standard sample preparations for target capture

Applications:

• FFPE samples
• Free-circulating DNA from plasma or serum
• Samples obtained by laser-capture microdissection of fresh, frozen or FFPE tissue
• Forensic samples
• Cells collected by flow cytometry
• Any other limiting or precious clinical sample

Kit Specifications and Contents / Storage:

Kits can be stored for up to 12 months at -20˚C.

Complete (Master Mix) kits include KAPA SYBR FAST qPCR Master Mix (2X), Primer Premix (41 bp, 129 bp and 305 bp, 10X) and a set of 5 DNA Standards. Primer Premixes and DNA Standards are also sold separately.

Components

Specifications