The Rapid DNA Lib Prep kit provides a streamlined and efficient method to construct DNA library for next generation sequencing (NGS) with Illumina (R) platforms. The entire protocol can be completed in 2 hours. Starting from fragmented input-DNA (1 - 1000 ng), the kit contains all the enzymes and buffers for end-preparation, adapter ligation and library amplification. To simplify the workflow, the fragmented DNA is end-repaired and adenylated during the initial step. 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 following workflow illustrates the processes of the Rapid DNA Lib Prep Kit.
|Tube name||8 RXN||24 RXN||96 RXN|
|End Preparation||End Prep Buffer||100 μl||260 μl||1000 μl|
|End Prep Enzymes||30 μl||78 μl||300 μl|
|Adaptor Ligation||Ligase MM||165 μl||429 μl||1650 μl|
|Ligase Mix||30 μl||78 μl||300 μl|
|Low-EDTA TE Buffer||1000 μl||2 mL||5 mL|
|Amplification||2X PCR Master Mix||250 μl||650 μl||2.5 mL|
|10X PCR Primers||40 μl||120 μl||500 μl|
The Low-EDTA TE buffer 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.
Required Materials Not Included
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)
PCR strip tubes or plates
AgencourtTM AMPure XP bead (Beckman Coulter Inc., cat. no. A63880)
Pipettes and multichannel pipettes
Aerosol resistant pipette tips
Agilent Bioanalyzer or comparable method to assess the quality of DNA library
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