Agilent SureSelect Target Enrichment System for SOLiD Fragment/Paired-End RNA-Seq Library Prep Protocol Version 1.1

About this manual

This guide describes recommended operational procedures to capture transcripts of interest using Agilent's SureSelect RNA Capture Kits and sample preparation kits for next-generation sequencing. The protocol is specifically developed and optimized to use Biotinylated RNA oligomer libraries, or Bait, to enrich targeted regions of the transcriptome.

The document details a comprehensive workflow including sample preparation (fragmentation, purification, reverse transcription), hybridization with capture probes, post-hybridization amplification, emulsion PCR, and library quantification using the Agilent 2100 Bioanalyzer. It includes specific instructions for required reagents, equipment, safety notes, and troubleshooting procedural steps.

What's inside

• Procedural Notes
• Safety Notes
• Required Reagents
• Sample Preparation
• Hybridization
• Post-Hybridization Amplification
• Reference

Frequently asked questions

What is the maximum target size for the Level MP4 SureSelect RNA Capture Kit?

The Level MP4 kit supports targets up to 3.0 Mb.

How long should you spin the tube after mixing frozen reagent stock solutions?

You must spin the tube in a centrifuge for 5 to 10 seconds.

What Biosafety Level rules should be followed when working with this system?

You must follow Biosafety Level 1 (BL1) safety rules.