The StepWise DNA Library Preparation Kit is designed to generate high-quality next generation sequencing (NGS) DNA libraries compatible with Illumina(R) platforms. Starting from fragmented input-DNA (500 pg - 1000 ng), the kit contains all the enzymes and buffers for end-repairing, dA-tailing, adapter ligation and library amplification. Due to the flexible protocol, the kit can be conveniently used for construction of PCR-free libraries from 100 ng of high-quality fragmented genomic DNA, or 10 ng of circulating cell-free DNA (cfDNA). PCR primers provided in the kit are optional for users who prefer self-selected high-fidelity DNA polymerases in the library amplification.
The Low-EDTA TE buffer and PEG/NaCl solution can be stored at room temperature or 4°C. All other components should be stored at -20°C. The shelf lives of all reagents are one year when stored properly.
Mechanical or enzymatic methods for DNA fragmentation Multiplex adapters (cat. no. RK20282, RK20283, RK20284, RK20285, RK20292, RK20293, RK20294, RK20295) compatible with Illumina(R) platforms 100% ethanol (ACS grade) Nuclease-free water PCR strip tubes or plates Magnetic stand Thermocycler AgencourtTM AMPure XP bead (Beckman Coulter Inc., cat. no. A63880) Pipettes and multichannel pipettes Aerosol resistant pipette tips Microcentrifuge Vortex mixer Agilent Bioanalyzer or comparable method to assess the quality of DNA library
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