Rapid Cell Free DNA Library Prep for Sequencing on Ion Torrent™ Platforms

The NEXTFLEX® DNA Sequencing Kit for Ion Torrent™ sequencing is designed to prepare genomic DNA-Seq or ChIP-Seq NGS libraries for sequencing using the Ion S5™, Ion S5™ XL, Ion Torrent™ PGM™, and Ion Proton™ sequencing platforms. The NEXTFLEX® DNA Sequencing Kit features Enhanced Adapter Ligation Technology, resulting in library preps with a larger number of unique sequencing reads. This DNA-Seq library prep kit simplifies the Ion Torrent™-based library prep workflow by using master mixed reagents and gel-based cleanup. In addition, the option of performing the reactions in a pre-prepared 96-well plate makes this kit more automation friendly than the other available versions.

Ion S5™, Ion PGM™ and Ion Proton™ DNA-Seq Multiplexing Options

The NEXTFLEX® DNA Barcodes for Ion Torrent™ sequencing are adapters containing indexed sequences that offer an improved multiplexing workflow and flexible setup. This format enables multiplexing of up to 64 samples. The NEXTFLEX® DNA Barcodes are available in sets of 8, 16, 32, and 64 unique adapters. The ability to pool samples in an efficient way significantly decreases hands-on time while providing robust data quality.

Features

• Enhanced Adapter Ligation Technology offers larger number of unique sequencing reads

• Flexible barcode options– Kits contain 8, 16, 32, or 64 unique barcodes

• Simplified workflow with 96-well plate compatible volumes

• Ideal for ChIP-seq and DNA-seq library construction for Ion Torrent™ sequencing

Kit Specs

NEXTflex™ DNA-Seq Kit Protocol

KIT CONTENTS

• NEXTFLEX® End Repair Buffer Mix

• NEXTFLEX® End Repair Enzyme Mix

• NEXTFLEX® Ligation Mix

• NEXTFLEX® Adapter Mix (6.25 µM)

• NEXTFLEX® Primer Mix (12.5 µM)

• NEXTFLEX® 5X PCR Master Mix

• 6X Loading Dye

• MW Ladder Ready-to-Load 100 bp

• Column Elution Buffer

• Nuclease-free Water

• Resuspension Buffer

• DNA Binding Buffer

• 5X DNA Wash Buffer

• Clean-Up Spin Columns

REQUIRED MATERIALS NOT PROVIDED

• 10 ng, 100 ng or 1 µg of fragmented genomic DNA in up to 40 µL nuclease-free water.

• NEXTFLEX® DNA Barcodes Ion Torrent Compatible – 8 / 16 / 32 / 64 (Cat # 4010-01, 4010-02, 4010-03, 4010-04)

• Ethanol 100% (room temperature)

• 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 (BioRad, Cat # MSB1001)

• Agencourt AMPure XP 5 mL (Beckman Coulter Genomics, Cat # A63880)

• Magnetic Stand -96 (Ambion, Cat # AM10027) / or / similar

• Heat block

• Thermocycler

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

• Nuclease-free barrier pipette tips

• Microcentrifuge

• 1.5 mL nuclease-free microcentrifuge tubes

• Low melt agarose such as Low Gelling Temperature Agarose with a melt point of 65ºC (Boston Bioproducts, Cat # P-730) / or / E-gel system

• 1X TAE buffer

• Clean razor or scalpel

• SYBR Gold (Invitrogen, Cat # S11494)

• UV transilluminator or gel documentation instrument

• Gel electrophoresis apparatus

• Electrophoresis power supply

• Vortex