Product nameDNA Library Preparation Kit (For Illumina®)
Assay time2h 30m
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 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.
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.
DNA Library Preparation Kit (For illumina)
PRICE UPON REQUEST
Store at -20°C. 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|
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab185903 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.