Rapid Plus DNA Lib Prep kit na 24 rxns ( NGS knihovny)

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.

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)
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