SEQuoia Complete Stranded RNA Library Prep Kit

Print
SEQuoia Complete Stranded RNA Library Prep Kit

Overview

Libraries prepared using the SEQuoia Complete Stranded RNA Library Prep Kit produce richer data sets. Libraries were prepared with either the SEQuoia Complete Stranded RNA Library Prep Kit or other commercial kits using 10 ng of rRNA-depleted RNA and sequenced to a read depth of 10 million reads.

A. Capture of mRNA

B. Capture of small RNA (<200 bases)

Co-detection of various nucleic acid targets in 5-plex reaction.
In addition to effectively capturing mRNA transcripts, libraries prepared with the SEQuoia Complete Stranded RNA Library Prep Kit included a far greater number of unique reads mapping to RNAs shorter than 200 nucleotides in length, including miRNA(), tRNA () snoRNA (), and snRNA (), indicating a richer, more complex library.
Better Library Yield with Fewer PCR Cycles

The SEQuoia Complete Stranded RNA Library Prep Kit produces substantially higher yields with fewer PCR cycles than other commercial kits when low quantities of degraded RNA are used as input. Ribosomal RNA was depleted using an NEBNext rRNA Depletion Kit1 from Frozen (A) and FFPE (B) Matched Pair Total RNA: Human Adult Normal Tissue: Liver2. Libraries were constructed using 100 ng or 10 ng of input with the SEQuoia Complete Stranded RNA Library Prep Kit (), NEBNext Ultra II Directional RNA Library Prep Kit (), or Illumina TruSeq Stranded Total RNA Library Prep Kit (). The total yields relative to the input RNA amount and number of PCR cycles are indicated.

A. RNA from frozen tissue

B. RNA from FFPE tissue

1 NEB #E6310
2 BioChain #R8234149-FP

Even Coverage Across Transcripts with Minimal Positional Bias

The SEQuoia RNA Library Prep Kit provides exceptional coverage, especially at the 5' end.
Human Placenta Total RNA (100 ng) was depleted of rRNA, used to prepare libraries, and then sequenced. The median coefficient of variation (CV) for coverage of the 1,000 most highly expressed transcripts is shown as a measure of transcript coverage. Relative to libraries produced using other commercial kits, the library produced using the SEQuoia Complete Stranded RNA Library Prep Kit produced a more uniform distribution of reads per transcript and had better coverage on the 5' end of transcripts.

Even Coverage Across Transcripts with Minimal Positional Bias

Library Prep Kit Coverage Uniformity (Median CV)
SEQuoia Complete Stranded RNA ()
0.46
NEBNext Ultra II Directional RNA ()
0.64
Illumina TruSeq Stranded Total RNA ()
0.66
Improved Coverage of GC-Rich Transcripts

The SEQuoia Complete Stranded RNA Library Prep Kit facilitates detection of GC-rich transcripts.
Coverage tracks for the JUND transcript, which has a GC content of 70%, are shown. Libraries prepared with the SEQuoia Complete Stranded RNA Library Prep Kit attain more even coverage (97%) than libraries prepared with the equivalent Illumina and NEB kits (91% and 94%, respectively).

Improved Coverage of GC-Rich Transcripts

The SEQuoia Complete Stranded RNA Library Prep Kit is a new RNA-Seq library prep kit with a proprietary enzyme designed to address the sensitivity and workflow limitations of current enzymes.
Read full description

Ordering Information


Description

Frequently Asked Questions

SEQuoia FAQ

Learn more about how to use the SEQuoia RNA Library Prep Kit.

View SEQuoia FAQs

The SEQuoia Complete Stranded RNA Library Prep Kit is a high-performance RNA-Seq kit that captures long and short RNAs in a single library, even from limited and low-quality samples. The streamlined workflow minimizes the number of pipetting steps and reduces the overall protocol time to less than 4 hours. The kit allows construction of libraries that are >99% stranded and suitable for next-generation sequencing on Illumina platforms and includes access to SEQSense Analysis Solution and Toolkit, a dedicated secondary data analysis pipeline.

Featuring SEQzyme, a proprietary engineered enzyme that couples cDNA synthesis with adapter addition in a continuous synthesis reaction. The unique enzymatic properties effectively captures all types and sizes of RNA species in a novel enzymatic reaction, significantly improving the diversity and quality of RNA libraries, even from limited or degraded RNA samples.

Experience the Power of SEQzyme

Features and Benefits

Highly Efficient and Streamlined Library Prep Workflow

The unique enzymatic properties of SEQzyme produce robust, high-quality RNA library. The kits are optimized to facilitate efficient library synthesis using a wide range of input RNA (100 pg to 1 μg) and are compatible with automated next-generation sequencing library construction. The streamlined workflow minimizes the number of pipetting steps and greatly reduces the overall protocol time to less than 4 hours.

Diverse Libraries with Low Quality and Limited Input Samples

The kit constructs a stranded (>99%) library that captures all RNA biotypes, both long and short RNAs, even from poor quality RNAs (for example, FFPE tissue samples). The feature enables researchers to discover biologically relevant differences in more areas of the transcriptome.

Greater diversity rna species

Dedicated Data Analysis Toolkit

SEQSense Analysis Solutions and Toolkit, which is available through a web portal, or can be downloaded and run locally, enables users to trim raw sequencing data and align sequences to a reference genome. The output provides specific quality control metrics to assess the quality of the sequencing data as well as a BAM file, which can be used with tertiary analysis packages to perform advanced analyses.

Applications and Uses

  • Differential gene expression and whole transcriptome profiling of high- and low-quality RNA samples
    • Sorted or single cell
    • Micro-dissection
    • Characterization of non-polyadenylated RNAs
    • Gene fusion identification
    • Single nucleotide variation discovery
  • Biomarker discovery
  • Splice junctions identification

Included Components

Each kit comes with 2 reagent boxes. Box A includes reagents for RNA fragmentation, cDNA synthesis, and library preparation. The kit is stored at –20°C:

  • Reagent A Fragmentation Mix
  • Reagent B End Repair Enzyme
  • Reagent C Poly(A) Buffer
  • Reagent D Poly(A) Polymerase
  • Reagent E SEQzyme Mix
  • Reagent F Amplification Mix

Box B includes magnetic bead for purification. The kit is stored at 4°C

SEQuoia Dual Indexed Primers are available and can be ordered separately.

Configuration Options

Catalog No. Description Size
17005726 SEQuoia Complete Stranded RNA Library Prep Kit 24 reactions
17005710 SEQuoia Complete Stranded RNA Library Prep Kit 96 reactions
12011928 SEQuoia Dual Indexed Primers Set, 12 vials of Unique Dual Indexes 96 reactions
12011930 SEQuoia Dual Indexed Primers Plate, one 96-well plate of Unique Dual Indexes 96 reactions

 

Specifications

Description Whole-transcriptome sequencing
Input Range 100 pg–1 µg RNA
Input Types High- and low-quality RNA samples, including degraded FFPE sample
Transcript Coverage >99% strandedness
Total Run Time <4 hours
Variant Class Gene fusions, novel transcripts, single nucleotide variation, transcript variants
System Compatibility Compatible with Illumina sequencing platforms
Product Configuration 24 or 96 reaction kit sizes
Shelf life Guaranteed 6 months from shipment when properly stored

Ordering

x

Account price failed to display. Please try reloading the page.Reload Page

If you are placing an order, you may proceed with your order; the account price will be applied if it is lower than the
list price. If the problem persists, please call customer service at or send an
email to

SEQuoia Complete Stranded RNA Library Prep Kit, 24 reactions

17005726
A high-performance RNA sequencing (RNA-Seq) kit that captures long and short RNAs in a single library, even from limited and low-quality samples. The kit provides all of the components required to make an RNA-Seq library for Illumina platforms. The size of the kit is for 24 reactions.

List Price: 
 
$1,803.00 
LoadingLoading...
EDU Price:
 
What is this?
Your Price: 
 
LoadingLoading...
Your Price:
 

Communication error

Reload Page

 
Quantity:   Add to Cart
 
SEQuoia Complete Stranded RNA Library Prep Kit, 96 reactions

17005710
A high-performance RNA sequencing (RNA-Seq) kit that captures long and short RNAs in a single library, even from limited and low-quality samples. The kit provides all of the components required to make an RNA-Seq library for Illumina platforms. The size of the kit is for 96 reactions

List Price: 
 
$6,695.00 
LoadingLoading...
EDU Price:
 
What is this?
Your Price: 
 
LoadingLoading...
Your Price:
 

Communication error

Reload Page

 
Quantity:   Add to Cart
 
SEQuoia Dual Indexed Primers Set

12011928
For use with SEQuoia Complete Stranded RNA Library Prep Kits. Contains 12 unique dual indexed primers supplied in 12 separate vials, for 96 reactions

List Price: 
 
$906.00 
LoadingLoading...
EDU Price:
 
What is this?
Your Price: 
 
LoadingLoading...
Your Price:
 

Communication error

Reload Page

Quantity:   Add to Cart
 
SEQuoia Dual Indexed Primers Plate

12011930
For use with SEQuoia Complete Stranded RNA Library Prep Kits. Contains 96 unique dual indexed primers plated in a 96-well plate, for 96 reactions

List Price: 
 
$906.00 
LoadingLoading...
EDU Price:
 
What is this?
Your Price: 
 
LoadingLoading...
Your Price:
 

Communication error

Reload Page

Quantity:   Add to Cart
 

Accessories

x

Account price failed to display. Please try reloading the page.Reload Page

If you are placing an order, you may proceed with your order; the account price will be applied if it is lower than the
list price. If the problem persists, please call customer service at or send an
email to

0.2 ml 8-Tube PCR Strips and Domed Cap Strips, high profile, clear

TBC0802
Pkg of 20, bag of 12 x 8 thin-wall polypropylene tube strips and 12 x 8-cap strips for PCR, natural (1,920 tubes and 1,920 caps)

List Price: 
 
$253.00 
LoadingLoading...
EDU Price:
 
What is this?
Your Price: 
 
LoadingLoading...
Your Price:
 

Communication error

Reload Page

Quantity:   Add to Cart
 
0.2 ml 12-Tube PCR Strips and Domed Cap Strips, high profile, clear

TBC1202
Pkg of 20, bag of 8 x 12 thin-wall polypropylene tube strips and 8 x 12-cap strips for PCR, natural (1,920 tubes and 1,920 caps)

List Price: 
 
$225.00 
LoadingLoading...
EDU Price:
 
What is this?
Your Price: 
 
LoadingLoading...
Your Price:
 

Communication error

Reload Page

Quantity:   Add to Cart
 

Documents

Number Description Options
7273 SEQuoia Complete Stranded RNA Library Prep Kit Product Information Sheet Click to download
10000119323 Instruction Manual, SEQuoia Complete Stranded RNA Library Prep Kit Click to download
10000119322 Instruction Manual, SEQuoia Dual Indexed Primers Click to download

This product contains electrical or electronic material. Learn more about Bio-Rad's EU Recycle Program

Get Help
Promotions
Sign Up for Bio-Rad Updates!
Enter your email address below to receive your choice of the latest news, promotions, and more.