KAPA hgDNA Quantification and QC Kit
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…
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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.
Features:
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
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
