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.

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

The NEXTFLEX^® DNA-Seq Library Prep Kits for Ion Torrent^™ sequencing contain the reagents to prepare 8 or 48 genomic or ChIP DNA samples for Ion Torrent^™ next-generation sequencing. The shelf life of all reagents is 12 months when stored properly at -20°C. Ships on dry ice.