SEQuoia Express Stranded RNA Library Prep Kit
SEQuoia Express Stranded RNA Library Prep Kit enables customers to overcome the challenge of a laborious library prep workflow by providing a cost-effective 3-hour, 3-tube, automation-friendly solution for constructing robust libraries that efficiently capture long RNA transcripts (>200bp).
Overview
SEQuoia Express Stranded RNA Library Prep Kit is a fast, high-performance RNA-Seq library preparation kit used to construct a sequence-ready library comprised of messenger RNA and long-noncoding RNA biotypes that is compatible with Illumina platforms from a broad range of total RNA inputs. Featuring SEQzyme, a proprietary engineered enzyme that couples cDNA synthesis with adapter addition in a continuous synthesis reaction, this kit streamlines library construction by reducing the number of components to just three tubes, significantly minimizing the number of pipetting steps and shortening the protocol time to under three hours.
SEQuoia Express Stranded RNA Library Prep Kit is compatible with SEQuoia RiboDepletion Kit for removal of constructs originating from ribosomal RNA and SEQuoia Dual-Indexed Primers, enabling a user to pool libraries and run them on a single flow-cell, greatly reducing the cost of over sequencing. Furthermore, it comes with a pre-packaged bioinformatics pipeline with the options to select a researcher’s local infrastructure or the cloud-based web application. This rapid and cost-effective workflow provides a seamless end-to-end solution for researchers who want to generate the highest quality data with minimal chance of error.
Features and Benefits
- Simplified workflow that can be completed under 3 hours
- Convenient 3-tube configuration minimizes pipetting steps and hands-on time
- Efficient and unbiased capture of coding and non-coding long RNA
- Validated with a broad range of RNA input
- Integrated with bioinformatics solution
Applications and Uses
For researchers and drug developers who require high-quality, complex, and unbiased sequencing libraries to profile differential expression patterns of mRNA and long non-coding RNA biotypes and be able to quickly assess the quality of their sequencing data. Main applications include:
- Liquid biopsy
- Single nucleotide variation discovery
- Gene fusion identification
- Splice junction identification
Included Components
Each kit comes with three reagent components: reaction mix, enzyme mix, and an amplification mix. The kit should be stored at -20°C.
SEQuoia Express Stranded RNA Library Prep Technology FAQs
The strandedness is maintained through the unique properties of SEQzyme, a highly engineered, multifunctional, patented retrotransposon. SEQzyme transcription is initiated on the 3’ end, therefore only 3’ ends of RNA will receive the 1st adapter. This highly efficient continuous synthesis reaction results in a sequencing-ready library with over 98% strandedness information preserved.
SEQuoia Express utilizes blunt end attachment of adapters to RNA fragments. The ratio of SPRI beads used for sample clean-up removes these adapter-dimers and other smaller RNA fragments. This results in the capture of mRNA and long non-coding RNAs (>200 bp).
SEQuoia Express is simplified by utilizing blunt end attachment of adapters to RNA fragments. This bypasses the ligation steps that involve many of the cumbersome pipetting and incubation steps required in other library prep kits and allows for a shorter, simpler workflow.
Unique molecular tags (UMI) are molecular tags typically of 6-12 random base pairs that allow researchers to distinguish transcript duplicates that originate from a single molecule. UMI is incorporated in our adapter design so researchers can easily identify PCR duplicates and detect bias that is introduced during the amplification process
Following library construction, assess the library quality on an Agilent Bioanalyzer 2100. The average peak size is approximately 350-450 bp.
100 ng Universal Human Reference RNA
100 ng Human Placenta RNA
SEQuoia Express Stranded RNA Library Prep Kit Information
The kit provides all reagents required to construct a library from RNA, with the exception of the SPRISelect beads from Beckman Coulter.
| Component | Cap Color | 24 Reactions (µl) | 96 Reactions µl | Storage Temperature | ||
| SEQuoia Express Reaction Mix | Violet | 600 | 2x1200 | -20°C | ||
| SEQuoia Express Enzyme Mix | Orange | 120 | 480 | |||
| SEQuoia Express Amplification Mix | Green | 600 | 2x1200 | |||
The kit is compatible with a broad dynamic range of input material, from 1 ng to 1 µg total RNA sample.
SEQuoia Express Stranded RNA Library Prep kit is guaranteed for 12 month shelf life after the shipping date if the product is stored properly at -20°C.
Yes! We offer two options of Unique Dual Indexed (UDI) Primers. They are pre-blended and balanced to achieve optimal multiplexed sequencing results. They are in 12 UDI or 96 UDI configurations. Visit bio-rad.com/SEQDIP for ordering information.
Yes! We suggest use SEQuoia Express Library Prep Kit with Bio-Rad’s multiplex-capable, post-amplification SEQuoia RiboDepletion Kit. Visit bio-rad.com/SEQuoiaRiboDepletion for ordering information.
SEQuoia Express Stranded RNA Library Prep Data Analysis
The difference relates to where the analysis takes place. The web hosted solution is a secure, cloud-based environment that provides an integrated preconfigured data analysis workflow that allows the user to perform initial analyses with only a few mouse clicks. The toolkit is a command line application packaged with required accessories, such as libraries and other dependencies. A user would download this docker container to run the application on their infrastructure.
We strongly recommend customer to use this complementary, customized data analysis software to process your sequencing data generated using SEQuoia library. For non-SeqSense user, you will require trimming of the contigs prior to alignment (guidelines are provided in the manual). In addition, it is likely that you will need to assign a reference library for non-gene RNA transcripts. For convenience, the SeqSense Analysis Software includes both of these functionalities.
Configuration Options
| Catalogue Number | Description | Size |
| 12017297 | SEQuoia Express Stranded RNA Library Prep Kit | 24 rxn |
| 12017265 | SEQuoia Express Stranded RNA Library Prep Kit | 96 rxn |
Related Products
- SEQuoia RiboDepletion Kit
- SEQuoia Dual Indexed Primers, Set
- SEQuoia Dual Indexed Primers, Plate
- CFX Opus Deep Well Real-Time PCR System
- 0.2 ml 8-Tube PCR Strips and Domed Cap Strips
- 0.2 ml 12-Tube PCR Strips and Domed Cap Strips
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Pkg of 1, kit for 24 reactions, includes 600 µl SEQuoia Express Reaction Mix, 120 µl SEQuoia Express Enzyme Mix, and 600 µl SEQuoia Express Amplification Mix
Pkg of 1, kit for 96 reactions, includes 2 x 1,200 µl SEQuoia Express Reaction Mix, 480 µl SEQuoia Express Enzyme Mix, and 2 x 1,200 µl SEQuoia Express Amplification Mix
Accessories
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)
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)
