PGN004 Sigma-AldrichPureGenome™ Next Generation Sequencing Library Validator Kit

This product has been discontinued.

Product Description

The quality of sequence obtained from Next Generation sequencing platforms such as Illumina, Inc. requires the accurate determination of input library DNA. Loading too much DNA can lead to overlapping clusters and loss of read counts whereas too little input can lead to poor read lengths and lower sequence output. The PureGenome™ NGS Library Validator Kit measures the absolute amount of adapter-ligated, sequence ready DNA in Illumina-compatible NGS libraries. In conjunction with a user-supplied SYBR^® Green Master Mix, the kit is a fast and reliable way to quantify DNA or cDNA sequencing libraries. The assay is capable of quantifying singleplex or multiplex (indexed) libraries via a straightforward qPCR test. This kit is optimized for use alongside the EMD Millipore Corporation PureGenome High Efficiency NGS Library Preparation Reagents: PureGenome NGS Library DNA Modifier (PGN001-1EA), PureGenome Library A-tail Enhancer (PGN002-1EA) and PureGenome NGS Library DNA Amplifier (PGN003-1EA), but is compatible with most DNA libraries designed for Illumina-based instruments.

*Note: PGN001-1EA, PGN002-1EA, PGN003-1EA should be purchased as a set for lot-matched qualtiy controlled solution for NGS library preparation.

Components

• 1 kit               PureGenome™ Next Generation Sequencing Library Validator Kit
  • 160 µl            PureGenome Validator Primer Mix (25X for up to 200 reactions)
  • 100 pM (20 µl)    PureGenome NGS DNA Control 1 (150 bp)
  • 100 pM (20 µl)    PureGenome NGS DNA Control 2 (300 bp)
  • 100 pM (20 µl)    PureGenome NGS DNA Control 3 (600 bp)

The kit contains a set of oligonucleotide primers developed for SYBR Green-based qPCR detection. The PureGenome Validator primers were designed to detect paired-end, adapter ligated and enriched DNA species with high amplification efficiency. The PureGenome NGS DNA Control standards are provided in three different sizes to more accurately match the representative library size. These standards are provided in 100 picomolar concentrations as a 10X stock along with primers at a 25X stock for the reaction mix. When following the respective protocol, this kit is sufficient for two hundred 20 µl reactions; however, this amount may vary based on user-specific needs and reagents (See TB575 for additional details).

Storage: Store all components at –20 °C. This kit is stable for at least one year at the recommended storage temperature.

Representative Data

See the figure at the bottom of this product page.