Next Gen Sequencing Kits 

Features

• Completely gel-free protocol with normal input amounts
• Greater discovery/detection rates reduce sequencing cost
• Randomized adapters reduce ligation bias, resulting in more accurate data
• Low input (1 ng) gel-based protocol included
• AIR™ Ligase offers greater sequencing depth
• Use more of your RNA sample – 10.5 µL input volume
• Barcoded primers are included in the kit allowing multiplexing of up to 48 samples
• An automation protocol is available for the PerkinElmer® Sciclone® G3 NGS/NGSx and the Zephyr® G3 NGS workstation
• Functionally validated with Illumina® sequencing platforms

Gel-Free Small RNA Library Prep with Randomized Adapters for Reduced Bias

The NEXTFLEX® Small RNA-Seq Kit v3 uses patented and patent-pending technology to provide a reduced-bias small RNA library preparation solution for Illumina® sequencing platforms with gel-free or low-input options. Our approach to reducing ligation-associated bias involves the use of adapters with randomized bases at the ligation junctions, resulting in greatly decreased bias in comparison to standard protocols. This reduction in bias results in data that more accurately represents abundances of small RNAs in the starting material. In addition, reduction of bias allows more miRNAs to be detected with fewer total reads, increasing efficiency and reducing cost for small RNA sequencing.

PAGE purification, required for traditional small RNA library prep, is tedious, time consuming, limits throughput, and prevents start-to-finish automation. The NEXTFLEX® Small RNA-Seq Kit v3 allows for gel-free small RNA library preparation. This is possible thanks to the dual approach used for adapter-dimer reduction. Unprecedented reduction of adapter-dimer formation allows completely gel-free small RNA library prep when starting with ≥200 ng of total RNA. Libraries prepared with the NEXTFLEX® Small RNA-Seq kit v3 have a higher proportion of reads mapping to miRNAs.

KIT CONTENTS

• NEXTFLEX® 3’ 4N Adenylated Adapter
• NEXTFLEX® 3’ Ligation Buffer
• NEXTFLEX® 3’ Ligation Enzyme Mix
• NEXTFLEX® Adapter Depletion Solution
• NEXTFLEX® Adapter Inactivation Buffer
• NEXTFLEX® Adapter Inactivation Enzyme
• NEXTFLEX® 5’ 4N Adapter
• NEXTFLEX® 5’ Ligation Buffer
• NEXTFLEX® 5’ Ligation Enzyme Mix
• M-MuLV Reverse Transcriptase
• NEXTFLEX® RT Buffer
• NEXTFLEX® Universal Primer
• NEXTFLEX® Barcode Primer
• NEXTFLEX® Small RNA PCR Master Mix
• 6X Loading Dye
• Ready to Load Low MW Ladder
• Resuspension Buffer
• Nuclease-free Water
• microRNA Control
• NEXTFLEX® Cleanup Beads
• NEXTFLEX® Elution Buffer

REQUIRED MATERIALS NOT PROVIDED

• 1 ng – 2 μg total RNA or purified small RNA from 1-10 μg total RNA in up to 10.5 μL
• Nuclease-free Water
• Isopropanol
• 80% Ethanol
• 2, 10, 20, 200 and 1000 μL pipettes
• RNase-free pipette tips
• Microcentrifuge
• 96 well PCR Plate Non-skirted
• Thin-wall nuclease-free PCR tubes
• Thermocycler
• Heat block
• Vortex
• Magnetic Stand -96