The Rapid Plus DNA Lib Prep kit provides a streamlined and efficient method to construction DNA library for next generation sequencing (NGS) at Illumina® 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 library 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 kit also provides DNA repair enzymes, which could repair DNA damages such as nicks, gaps, oxidized bases, damaged/blocked 3’ ends, AP sites (apurinic/apyrimidinic sites), and uracil bases. In addition, the DNA repair enzymes significantly increase the NGS library yields from FFPE-derived DNA.
The following workflow illustrates the processes of Rapid DNA Lib Prep Kit.
|Tube name||8 RXN||24 RXN||96 RXN|
|End Preparation||End Prep Buffer||70 μl||168 μl||700 μl|
|End Prep Enzymes||30 μl||78 μl||300 μl|
|Damaged DNA Repair Enzymes||30 μl||78 μl||300 μl|
|Adaptor Ligation||Ligation Buffer||300 μl||780 μl||3 ml|
|Ligation Enzymes||100 μl||240 μl||1000 μl|
|Amplification||2X PCR Master Mix||250 μl||650 μl||2.5 mL|
|10X PCR Primers||50 μl||130 μl||500 μl|
All 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® 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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