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AMPINEXT™ DNA Library Preparation Kit (Illumina)

ENZ-GEN504

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Complete set of optimized reagents to carry out a successful DNA library preparation.

Fast and streamlined procedure – The procedure from fragmented DNA to size selection is less than 2 h 30 min.
Highly Convenient – The kit contains all required components for each step of DNA library preparation.
Minimized bias – Ultra HiFi amplification and an optional PCR-free step.
Flexibility – Use for both non-barcoded (singleplexed) and barcoded (multiplexed) DNA library preparation.

The AMPINEXT™ DNA Library Preparation Kit (Illumina) is a complete set of optimized reagents to carry out a successful DNA library preparation. The kit is suitable for preparing a DNA library for next generation sequencing applications using an Illumina sequencer, which includes genomic DNA-seq, ChIP-seq, MeDIP/hMeDIP-seq, 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 DNA Library Preparation Kit (Illumina) Schematic — Figure 1. Schematic Procedure for using the AMPINEXT™ DNA Library Preparation Kit (Illumina)

Product Details

Application Notes	A complete set of optimized reagents to carry out a successful DNA library preparation.
Assay Time	2 hours 30 minutes
Contents	10X End Repair Buffer End Repair Enzyme Mix 10X dA-Tailing Buffer Klenow Fragment (3’-5’ exo^–) 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 ChIP reactions, MeDIP/hMeDIP reaction, or exon capture. DNA should be relatively free of RNA since large fractions of RNA will impair end repair and dA tailing, resulting in reduced ligation capabilities. Input amount of DNA can be from 5 ng to 1 µg. For optimal preparation, the input amount should be 100 ng to 200 ng. For amplification-free, 500 ng or more is needed.

Handling & Storage

Use/Stability	Upon receipt: Store the following components at -20°C immediately: 10X End Repair Buffer, End Repair Enzyme Mix,10X dA-Tailing Buffer, Klenow Fragment (3’-5’ exo-), 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 to carry out a successful DNA library preparation. In the library preparation, DNA is first fragmented to the appropriate size (about 300 bp peak size). The end repair of the DNA fragments is performed and an A-overhang is added at the 3'-end of each strand. Adaptors are then ligated to both ends of the end repaired/dA tailed DNA fragments for amplification and sequencing. Fragments are then size selected and purified with MQ beads, which allows for quick and precise size selection of DNA. Size-selected DNA fragments are then amplified with a high-fidelity PCR Mix which 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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