NEXTFLEX® Rapid DNA-Seq Kit 2.0 for Illumina® Platforms

The NEXTFLEX® rapid DNA-seq 2.0 kit is designed to prepare multiplexed libraries for single or paired-end sequencing using Illumina® platforms. The enhanced NEXTFLEX® kit simplifies workflow by using master mixed reagents and magnetic bead based cleanup, reducing pipetting and eliminating time-consuming steps in library preparation. The NEXTFLEX® rapid DNA-seq kit 2.0 produces library preps with a larger number of unique sequencing reads, allowing for robust genome coverage in challenging GC-rich sequences that can introduce bias.

The NEXTFLEX® rapid DNA-seq 2.0 kit bundle offers all the advantages and reagents of the NEXTFLEX Rapid DNA-seq kit 2.0 and includes  NEXTFLEX-HT™ barcodes (8, 48, or 96 reaction size kits) supplied in a single-use format. This allows the lab to tie all the necessary components of the library prep workflow to a single lot for ease of purchasing and tracking of consumables.

Features

• Complete library prep solution, including size selection beads

• Bundle kits available with library prep reagents and barcodes

• Up to 384 barcodes available upon request

• Convenient and cost-effective package in a 8, 48, or 96 reaction format

• Simplified workflow

• Automation compatible with liquid handlers, including the PerkinElmer Sciclone® G3 NGS/NGSx workstations and Zephyr® G3 NGS workstation

• Robust genome coverage

• Reliable performance with GC-bias and sequencing bias mitigation

• Low input requirement for genomic DNA down to 1 ng

• For FFPE samples, use the NEXTFLEX® Rapid XP DNA-Seq Kit

• Functionally validated with Illumina® sequencing platforms

Kit Specs

Protocol

Figure 1: Libraries prepared with NEXTFLEX® rapid DNA-Seq kit 2.0 show higher yield than libraries prepared with Competitor K’s kit using the same number of PCR cycles from a 250 ng and 1 ng input (A) and robust conversion rate (B).

Additional Data between NEXTFLEX® Rapid DNA-Seq 2.0 and Competitor K’s Kit Libraries

Figure 2: Libraries prepared with NEXTFLEX® Rapid DNA-seq 2.0 shows less GC bias than libraries prepared with Competitor K’s kit.

Figure 3: Libraries prepared with NEXTFLEX® Rapid DNA-seq 2.0 shows higher genome coverage than libraries prepared with Competitor K’s kit.

KIT CONTENTS

• NEXTFLEX® End Repair & Adenylation Buffer Mix 2.0

• NEXTFLEX® End Repair & Adenylation Enzyme Mix 2.0

• NEXTFLEX® Ligase Buffer Mix 2.0

• NEXTFLEX® Ligase Enzyme 2.0

• NEXTFLEX® PCR Master Mix 2.0

• NEXTFLEX® Primer Mix 2.0

• NEXTFLEX® Cleanup Beads 2.0

• Nuclease-free Water

• Resuspension Buffer

REQUIRED MATERIALS NOT PROVIDED

• 1 ng – 1 µg of fragmented genomic DNA in up to 32 µL nuclease-free water.

• NEXTflex® barcodes if purchasing a non-bundled kit (includes NEXTflex-HT™ barcodes)

• Ethanol 100% (room temperature)

• Ethanol 80% (room temperature)

• Covaris System (S2, E210) or other method for DNA fragmentation

• 96 well PCR Plate Non-skirted (Phenix Research, Cat # MPS-499) or similar

• 96 well Library Storage and Pooling Plate (Thermo Fisher Scientific, Cat # AB-0765) or similar

• Adhesive PCR Plate Seal (Bio-Rad, Cat # MSB1001)

• Magnetic Stand -96 (Thermo Fisher Scientific, Cat # AM10027) or similar

• Heat block

• Thermocycler

• 2, 10, 20, 200 and 1000 µL pipettes / multichannel pipettes

• Nuclease-free barrier pipette tips

• Microcentrifuge

• 1.5 mL nuclease-free microcentrifuge tubes

• Vortex

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