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AMPINEXT™ High-Sensitivity DNA Library Preparation Kit (Illumina)

ENZ-GEN505

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A complete set of optimized reagents to prepare a DNA library from very small amount of samples for use in next-generation sequencing applications.

High sensitivity and flexibility – Use with non-barcoded (singleplexed) and barcoded (multiplexed) DNA library preparation.
Fast and streamlined procedure – No clean-up is required between each step and all reactions take place in the same tube.
Highly convenient – The kit contains all required components for each step of DNA library preparation,
Minimized bias – Ultra HiFi amplification and optional PCR-free step.

The AMPINEXT™ High-Sensitivity DNA Library Preparation Kit (Illumina) is a complete set of optimized reagents to prepare a DNA library from very small amount of samples for use in next-generation sequencing applications. This kit is suitable for preparing a DNA library using sub-nanogram amounts of DNA input for next generation sequencing applications using an Illumina sequencer. These applications include genomic DNA-seq, ChIP-seq, MeDIP/hMeDIP-seq, traditional bisulfite-seq, and targeted re-sequencing. The optimized protocol and components of the kit allow both non-barcoded (singleplexed) and barcoded (multiplexed) DNA libraries to be constructed quickly with reduced bias.

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AMPINEXT High-Sensitivity DNA Library Preparation Kit (Illumina) schematic — Figure 1. Schematic Procedure for using the AMPINEXT High-Sensitivity DNA Library Preparation Kit

Product Details

Application Notes	A complete set of optimized reagents to prepare a DNA library from very small amount of samples for use in next-generation sequencing applications.
Assay Time	1 hour 30 minutes
Contents	10X End Polishing Buffer End Polishing Enzyme Mix End Polishing Enhancer 2X Ligation Buffer T4 DNA Ligase Adaptors (50 µM) MQ Binding Beads 2X HiFi PCR Master Mix Primer U (10 µM) Primer I (10 µM) Elution Buffer
Technical Info / Product Notes	Starting materials can include fragmented dsDNA isolated from various tissue or cell samples, dsDNA enriched from a ChIP reaction, MeDIP/hMeDIP reaction, or exon capture. DNA should be relatively free of RNA because large fractions of RNA will impair end repair and dA-tailing, resulting in reduced ligation capabilities. The input amount of DNA can be from 0.2 ng to 100 ng. For optimal preparation, the input amount should be 10 ng to 50 ng.

Handling & Storage

Use/Stability	Upon receipt: Store the following components at -20°C immediately: 10X End Polishing Buffer, End Polishing Enzyme Mix, End Polishing Enhancer, 2X Ligation Buffer, T4 DNA Ligase, Adaptors, 2X HiFi PCR Master Mix, Primer U, Primer I, and Elution Buffer. Store the following components at 4°C: MQ Binding Beads. Store all other components at room temperature.
Shipping	Blue Ice

Regulatory Status	RUO – Research Use Only

Datasheet, Manuals, SDS & CofA

Manuals And Inserts

Specific Protocol

This kit includes all reagents required at each step of the workflow to carry out a successful DNA library preparation. In the library preparation, DNA is first fragmented to an appropriate size (about 300 bps in peak size). The end repair/dA tailing (end polishing) of the DNA fragments are performed simultaneously. Adaptors are then ligated to both ends of the polished DNA fragments for amplification and sequencing. Ligated fragments are size selected and purified with MQ beads, which allows for quick and precise size selection of DNA. Size-selected DNA fragments are amplified with a high-fidelity PCR Mix that ensures maximum yields from minimum amounts of starting material and provides highly accurate amplification of library DNA with low error rates and minimum bias.

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