DNA Library Preparation Kit (For Illumina®) (ab185903)
Key features and details
- Assay time: 2 hr 30 min
- Sample type: DNA
Overview
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Product name
DNA Library Preparation Kit (For Illumina®) -
Sample type
DNA -
Assay time
2h 30m -
Product overview
The DNA Library Preparation Kit (For Illumina®) (ab185903) 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.
Starting Materials
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.Illumina® is a registered trademark of Illumina, Inc.
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Notes
DNA library preparation is a critical step for next generation sequencing (NGS). To generate accurate sequencing data for NGS, the prepared library DNA should be sufficient in yield and of high quality. Also, as NGS technology is continuously improving, DNA library preparation is required to be optimized accordingly. Most of the currently used methods are unfortunately time-consuming, expensive, and inconvenient. Some of the methods are relatively quick by combining end repair and dA tailing or even ligation in one-step, but have been shown to generate significant G tailing or form concatmers at the ligation step or have high insertion bias. These side reactions eventually result in the prepared DNA library being less efficient and inaccurate. An ideal DNA library preparation method should be balanced in speed, convenience, small sample-suitability, cost-effectiveness, and accuracy.
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Tested applications
Suitable for: ChIP-sequencingmore details
Properties
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Storage instructions
Please refer to protocols. -
Components 12 tests 24 tests 10X dA-Tailing Buffer 1 x 40µl 1 x 80µl 10X End Repair Buffer 1 x 40µl 1 x 80µl 2X HiFi PCR Master Mix 1 x 160µl 1 x 320µl 2X Ligation Buffer 1 x 250µl 1 x 500µl Adaptors (50 μM) 1 x 15µl 1 x 30µl Elution Buffer 1 x 1ml 1 x 2ml End Repair Enzyme Mix 1 x 25µl 1 x 50µl Klenow Fragment (3’-5’ exo-) 1 x 15µl 1 x 30µl MQ Binding Beads 1 x 1.6ml 1 x 3.2ml Primer I (10 μM) 1 x 15µl 1 x 30µl Primer U (10 μM) 1 x 15µl 1 x 30µl T4 DNA Ligase 1 x 15µl 1 x 30µl -
Research areas
Images
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Size distribution of library fragments
Human placenta DNA was sheared to 210 bps peak size and 20 ng of sheared DNA was used for DNA library preparation.