NEXTFLEX Cystic Fibrosis Amplicon Panel2018-08-31T19:07:34+00:00

VALIDATED PANEL FOR DETECTION OF THE CFTR GENE WITH LIBRARY PREP REAGENTS AND BARCODES FOR ILLUMINA® SEQUENCING

NEXTFLEX® Cystic Fibrosis Amplicon Panel

  • 100% coverage of CFTR exons and flanking intron-exon boundaries
  • 99% uniformity of amplicons at 0.2x coverage and 100% uniformity of amplicons at 0.1x coverage
  • 384 unique barcodes available upon request
  • Complete solution for targeted sequencing, including gene specific primers, PCR Master Mix, clean-up beads, and barcodes
  • Low input – Only 20 ng DNA isolated from fresh or frozen samples required (2.5 ng input optimized protocol available)

  • NOVA-4231-01

       $386

    8 BACODES | 8 RXNS

  • NOVA-4231-02

       $2,163

    48 BACODES | 48 RXNS
Download Manual
Covered Regions
For research use only. Not for use in diagnostic procedures.

Coverage of the CFTR Gene for Illumina® Sequencing

The NEXTFLEX® Cystic Fibrosis Amplicon Panel allows you to quickly target and sequence 28 exons of the CFTR locus. This kit contains primer pairs and reagents needed to amplify these coding regions of the CFTR gene from DNA isolated from fresh or frozen material, and prepare libraries for Illumina® sequencing.

The NEXTFLEX® Cystic Fibrosis Amplicon Panel, the panel has the capability of multiplexing up to 375 samples at 100x coverage for detection of germline mutations on a single Illumina® 2×250 MiSeq® lane.

A Complete Solution for Targeted CFTR Sequencing

The NEXTFLEX® Cystic Fibrosis Amplicon Panel is a complete, validated solution that targets 28 exons in the CFTR locus using 61 PCR primer pairs. This kit requires only 20 ng of DNA isolated from fresh or frozen samples (optional 2.5 ng input protocol available). The convenient predesigned panel allows researchers to focus on data generation and analysis, not on labor-intensive primer design and target selection. This kit can be easily automated on liquid handlers, further simplifying workflow.

KIT CONTENTS

  • NEXTFLEX® CFTR Amplicon Primer Mix 1
  • NEXTFLEX® CFTR Amplicon Primer Mix 2
  • NEXTFLEX® Hot Start PCR I Master Mix
  • NEXTFLEX® Stop Solution
  • NEXTFLEX® Ligation Mix
  • NEXTFLEX® Amplicon DNA Adapter
  • NEXTFLEX® PCR II Barcoded Primer Mix
  • NEXTFLEX® PCR II Master Mix
  • Nuclease-free Water
  • Resuspension Buffer
  • NEXTFLEX® Cleanup Beads

REQUIRED MATERIALS NOT PROVIDED

  • 20 – 100 ng of extracted genomic DNA (two 10 – 50 ng aliquots in up to 19.2 μL nuclease-free water each)
  • Ethanol 80% (room temperature)
  • 96 well PCR Plate Non-skirted (Phenix Research, Cat # MPS-499) or similar
  • 96 well Library Storage and Pooling Plate (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
  • 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

Amplicon Panel Flow Chart

The NEXTFLEX®  Cystic Fibrosis Amplicon Panel contains enough material to prepare 8, 48, or 96 samples for Illumina® sequencing from DNA isolated from fresh or frozen samples. The shelf life of all reagents is 12 months when stored properly. All components can be safely stored at -20°C.

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