Productos - NGS, Librerias y Servicios - Librerías - RNA Sequencing

Zymo-Seq SwitchFree 3′ mRNA Library Kit

Ref. R3008

HIGHLIGHTS

Simplest protocol: RNA to library in less time with easy handling driven by the SwitchFree™ technology.
High performance: Built-in Unique Molecular Identifiers (UMIs) allow for accurate deduplication maximizing unique reads.
Low input compatible: Utilize as little as 10 ng total RNA without prior mRNA enrichment.

DESCRIPTION

Zymo-Seq SwitchFree 3′ mRNA Library Kit is an all-in-one 3′ mRNA library prep kit that provides a simple workflow to generate mRNA libraries for next-generation sequencing in about 4 hours. The kit’s simplicity is driven by the novel SwitchFree technology that ligates P7 adapter directly to the first-strand cDNA without “switching” to a different strand.

This simplifies reagent handling for easy and fast library preparations. This Zymo-Seq SwitchFree 3′ mRNA Library Kit features built-in unique molecular identifiers (UMIs) allowing accurate evaluation of duplication rate and quantification of gene expression. This all-in-one 3′ mRNA library prep kit is compatible with an input as little as 10 ng of total RNA, making it an ideal tool for simple and efficient mRNA expression analysis. This kit offers scalable sample multiplexing suitable for high-throughput gene counting.

The Zymo-Seq SwitchFree 3′ mRNA Library Kit contains all the necessary reagents required for 3′ mRNA library preparation, including unique dual indexing primers and the magnetic beads for cleanup.

TECHNICAL SPECIFICATIONS

Equipment Needed (user provided)	Thermal cycler with heated lid, magnetic stand for 0.2 mL PCR tubes, and microcentrifuge for 0.2 mL PCR tubes and 1.5 mL microcentrifuge tubes.
Input Quality	RNA should be free of DNA contamination and enzymatic inhibitors, with A260/A280 and A260/A230 ≥ 1.8. RNA with lower purity ratios (A260/A280 and A260/A230) should be treated with DNase I and purified with the RNA Clean & Concentrator™ (Cat. No. R1013) prior to processing. RNA should be suspended in water, TE, or a low-salt buffer. Intact RNA (RNA Integrity Number or RIN ≥ 8.0) performs optimally with this kit. For degraded RNA input, e.g., those with a size distribution peaking at ≤ 500 nt, or RIN < 4, see Section 1, Step 4 of the protocol for the suggested modification. Additionally, use a higher amount of input whenever possible.
Library Storage	Libraries eluted in DNA Elution Buffer (provided) may be stored at ≤ 4°C overnight or ≤ -20°C for long-term storage.
Processing Time	~4 hours
RNA Input	Compatible with 10 – 500 ng of total RNA.
Sequencing Platform Compatibility	Libraries are compatible with all Illumina^® sequencing platforms except HiSeq^® X. (Illumina^® originally limits the applications on HiSeq^® X exclusively for whole-genome libraries. Please confirm with the sequencing service provider for acceptability and additional details if expecting to sequence Zymo-Seq SwitchFree™ 3′ mRNA libraries on HiSeq^® X Series sequencers.)