Preparation of library DNA is an important process for successful cluster formation on Illumina® platforms which is required to generate valuable sequencing data. This kit provides room temperature stable reagents to convert double stranded fragmented DNA into libraries. The SPARK™ DNA Sample Prep Kit is primarily designed for constructing genomic DNA libraries, paired-end DNA libraries, or paired-end multiplexed DNA libraries. All enzymes and buffers required are provided as room temperature stable mixes that can convert 10 ng to 1 µg of fragmented dsDNA into library DNA.
DNA Library Construction Assay
DNA fragmented at 400 to 500 base pairs was blunt ended using re-hydrated End Repair Mix. Deoxyadenosine was added to the 3′ ends using re-hydrated A-Tailing Mix and ligated to double-stranded barcoded DNA adaptors using re-hydrated Ligation Mix. The adaptor ligated DNA was then subjected to PCR amplification and samples from all steps were analyzed using a HS DNA chip on an Agilent Bioanalyzer.
End Repair Mix (LY0001V-1): A DNA polymerase produces blunt ended dsDNA fragments and a DNA kinase phosphorylates their 5′ ends.
A-Tailing Mix (LY0002V-1): A DNA polymerase adds a single deoxyadenosine monophosphate residue to the 3′ end of blunt ended dsDNA fragments.
Ligation Mix (LY0003V-1): A DNA Ligase installs barcoded dsDNA adapters with a deoxythymidine monophosphate overhang to dsDNA fragments with 3′ dAMP overhangs.
Kit enzyme components were tested prior to assembly and found free of contaminating endonucleases and exonucleases. Enzyme purity was >99% as determined by SDS-PAGE and negligible E.coli genomic DNA contamination was confirmed by qPCR. Specific activity was verified for each enzyme pre and post-lyophilization.
|SPARK™ DNA Sample Prep Kit for Illumina®|
|Unit Size||8 Reactions|
|SDS||Available on request|
|Assay||DNA Library Construction Assay|
* For a detailed summary of assay conditions and data, refer to the Quality Controls Analysis section.
Limitations of Use
This product was developed, manufactured, and sold for in vitro use only. The product is not suitable for administration to humans or animals. MSDS sheets relevant to this product are available upon request.
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