Custom Premix Selecting "Custom Premix" option means that all of the beads you have chosen will be premixed in manufacturing before the kit is sent to you.
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.
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.
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.