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For labs that routinely run DNA sequencing, Revvity® has developed the NEXTFLEX® rapid DNA-seq kit 2.0 that boasts high yields of quality sequencing libraries from fragmented genomic DNA in the form of a convenient, all-inclusive kit that includes the library prep reagents, barcoded adapters, and cleanup beads. Labs have the option of multiplexing up to 384 samples using the NEXTFLEX-HT™ barcodes or up to 192 samples using the NEXTFLEX® unique dual index barcodes.
Brand: Revvity
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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 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 core kit, but with the additional benefit of including unique NEXTflex-HT™ barcodes for the number of reactions (8, 48, or 96 reaction size kits) 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.
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).
The NEXTflex® adapters are long, annealed adapters containing indexed sequences, which offer an improved multiplexing workflow and flexible experimental setups catered to individual needs. Our barcode portfolio encompass options for single or paired-end multiplex sequencing, and the NEXTflex® rapid DNA-seq kit 2.0 is compatible with the NEXTflex® DNA Barcodes and NEXTflex-96™ DNA Barcodes, NEXTflex® ChIP-Seq Barcodes, and NEXTflex-96™ ChIP-Seq Barcodes. The NEXTflex®rapid DNA-seq kit 2.0 reagents are also compatible with the NEXTflex® Unique Dual Index Barcodes for up to 192 sample multiplexing.
The NEXTflex® rapid DNA-seq kit 2.0 bundles come provided with the NEXTflex-HT™ Barcodes.
The NEXTflex® rapid DNA-seq kit 2.0 is compatible with multiple liquid handlers, including the Revvity® Sciclone® G3 NGS workstation.
For larger volume requirements, customization and bulk packaging is available.
Size selection is a critical issue in NGS library preparation. Our blog post, Tech Tips – When to Size Select, addresses a number of factors to consider before determining a size selection strategy.
Kit Specs
The NEXTflex® rapid DNA-seq 2.0 kits contain library prep reagents, including cleanup beads, to prepare 8, 48, or 96 genomic DNA samples for Illumina® sequencing. The bundled solution also includes the corresponding number of unique NEXTflex-HT™ barcodes per reaction. The shelf life of all reagents is 6 months when stored properly at -20°C. This kit is shipped on dry ice.
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
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
Figure 1: Libraries prepared with NEXTflex® Rapid DNA-seq 2.0 shows less GC bias than libraries prepared with Competitor K's kit.
Figure 2: Libraries prepared with NEXTflex® Rapid DNA-seq 2.0 shows higher genome coverage than libraries prepared with Competitor K's kit.
What tools you should use to analyze your DNA-Seq data?
Read our blog post, Analysis Recommendations for DNA-Seq, to find out what tools we recommend to perform secondary and tertiary analysis.
