Next Gen Sequencing Kits 

Features

• Combined library prep workflow for both small RNA and mRNA sequencing
• Completely gel-free protocol from inputs as low as 5 ng
• Compatible with total RNA inputs, no poly(A) selection or rRNA depletion required
• Complete kit solution, including cleanup/size selection beads and barcodes*
• Automated on the PerkinElmer Sciclone® G3 NGS/NGSx workstations

Gel-free, Combined Small RNA and mRNA Library Prep with Randomized Adapters to Reduce Ligation Bias and UDIs to Prevent Sample Mis-assignment on Illumina® Platforms

The NEXTFLEX® Combo-Seq™ Kit enables the user to generate combined mRNA and small RNA libraries in a single workflow using 5 ng – 100 ng of total RNA inputs, without requiring upfront costly and tedious rRNA depletion or poly(A) selection. The kit utilizes patented and patent-pending technology that allows processing of mRNA fragments and small RNA for library construction. Like the best-in-class, NEXTFLEX® small RNA-seq kit v3, the NEXTFLEX® Combo-Seq™ mRNA/miRNA library preparation kit utilizes adapters with randomized ends to greatly reduce bias compared to standard protocols. This allows a more accurate representation of small RNA in the starting material and cost-saving due to efficient sequencing. Adapter-dimer reducing techniques built into the protocol enables gel-free workflow from inputs as low as 5 ng of total RNA. As a result, the kit is streamlined and Automation-friendly from start-to-finish. Libraries with robust yield can be prepared within one day using a manual or automated protocol.

KIT CONTENTS

• NEXTFLEX® Anchored Oligo(dT) Primer
• NEXTFLEX® RT/RNase H Annealing Buffer
• NEXTFLEX® RT/RNase H Reaction Buffer
• NEXTFLEX® Combo-Seq™ Reverse Transcriptase
• NEXTFLEX® Thermostable RNase H
• NEXTFLEX® PAP Buffer
• NEXTFLEX® Poly(A) Polymerase
• NEXTFLEX® tRNA/YRNA Blocker
• Adapter Depletion Solution
• NEXTFLEX® Combo-Seq™ 5’ Ligation Buffer
• NEXTFLEX® Combo-Seq™ 5’ 4N Adapter
• NEXTFLEX® 5’ Ligation Enzyme Mix
• NEXTFLEX® PAP Buffer
• NEXTFLEX® Poly(A) Polymerase
• NEXTFLEX® Combo-Seq™ RT Buffer
• NEXTFLEX® PAP Buffer
• NEXTFLEX® UDI Barcoded Primer Mix
• NEXTFLEX® PCR Master Mix
• Nuclease-free Water
• Resuspension Buffer
• NEXTFLEX® Cleanup Berads

REQUIRED MATERIALS NOT PROVIDED

• 5 ng – 100 ng total RNA in up to 11 μL Nuclease-free Water
• Human Universal RNA or other total RNA, as a control (optional)
• Isopropanol
• 80% Ethanol, freshly prepared
• 10, 20, and 200 μL pipettes
• 10, 50, and 200 μL multichannel pipettes
• RNase-free barrier pipette tips
• 96 well PCR Plate Non-skirted
• Nuclease-free PCR strip tubes with caps 
• Nuclease-free 1.7 mL microcentrifuge tubes
• Thermal cycler
• Vortex
• Microcentrifuge
• Magnetic Stand -96