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Low Input NGS Library Prep

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The PureGenome™ Low Input NGS Library Construction Kit streamlines the process of generating indexed, or barcoded libraries, for Illumina® HiSeq® sequencing from extremely low amounts of input DNA (from 50 pg to 1 ng). This kit is well suited for applications such as ChIP-Seq (chromatin immunoprecipitation followed by next generation sequencing) as well as other sequencing applications where input DNA is limited.

To achieve this level of sensitivity, the PureGenome™ Low Input NGS kit utilizes a unique technology that adds adaptors to sheared genomic DNA with ultra-high efficiency. This methodology enables the previous limits of NGS library construction to be reduced to below 1 ng. Additionally, this approach greatly simplifies NGS library construction. As compared to conventional approaches that require DNA isolation between protocol steps, the PureGenome™ Low Input NGS process occurs in a single tube and does not require any in process DNA isolation steps. Once library construction is complete, in a final step, DNA purification and size selection is performed using the Agencourt® AMPure® XP beads provided with this kit.
  • Sensitive: Generate libraries with as little as 50 pg of input DNA 
  • Fast: Construct instrument ready libraries in about 2 hours
  • Convenient: Single tube repair, ligation, and amplification steps & no DNA purification step required.
  • Robust: High diversity, low duplicate reads. 
  • Proven: Demonstrated performance in ChIP-seq library construction 
  • Reproducible: Get reproducible libraries across 50pg – 50ng range
  • Complete: All key reagents provided including 12 Illumina indexing barcodes and Agencourt® AMPure® XP beads for recovery and concentration of final library.

PureGenome low input NGS Kit Library Construction Workflow
Starting with as little as 50 pg of fragmented double-stranded DNA or cDNA, PureGenome™ Low Input NGS kit creates indexed libraries in 3 simple steps: end repair, adapter ligation, and high-fidelity library amplification. The streamlined workflow prevents sample loss and is performed in about 2 hours in a single tube or well preventing sample loss and enhancing positive sample identification.

PureGenome low input NGS Kit ChIP-Seq Data
Data shows that REST associates strongly with the NPAS4 gene, the expression of which it is known to repress. Even a small amount of input DNA (50 pg) showed a strong signal when using PureGenome™ Low Input NGS Library Construction Kit, as compared to 1 ng DNA with a standard protocol. Chromatin immunoprecipitation was performed using the Magna ChIP® HiSens kit (17-10460), anti-REST antibody (2 μg of cat# 09-019), 20 μL Protein A/G beads, and 4x106 cross-linked HeLa cell chromatin followed by DNA purification using magnetic beads. Libraries were prepared from Input and ChIP DNA samples using PureGenome™ Low Input NGS Library Construction Kit (from 50 pg and 300 pg of ChIP DNA) or standard protocol (NGS library construction kit obtained from Supplier N using 1 ng of ChIP DNA) with Illumina barcoded adapters. In all cases constructed libraries were analyzed on an Illumina HiSeq™ instrument.

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