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SeekGene SeekOne DD scFAST-seq kit

Waren-Nr. PST2
8 Tests/Kit
Full-length RNA Transcriptome

Product Information

Product Overview

SeekOne® Digital Droplet (SeekOne® DD) High throughput Single Cell Full-length RNA Sequence Transcriptome-seq (scFAST-seq) Kit, selfdeveloped by Beijing SeekGene BioSciences Co., Ltd., is a powerful commercial tool for high-throughput whole transcriptome profiling. The scFAST-seq method makes use of innovative techniques including semi-random primers, efficient reverse transcription, template swapping, and effective rRNA removal to build full-length RNA libraries of up to 12,000 cells.

Compared to conventional 3' scRNA-seq, scFASTseq has distinct advantages in detecting nonpolyadenylated transcripts, transcript coverage length, and identification of more splice junctions. With target region enrichment, scFAST-seq can simultaneously detect somatic mutations and cell states in individual tumour cells, providing valuable information for precision medicine.

The kit is designed to be used in conjunction with our SeekOne® Digital Droplet System (SeekOne® DD) to complete the entire process from single-cell nucleic acid labelling to transcriptome library construction. When equipped with "SeekSoul Tools", our single-cell data analysis software, we provide a one-stop-shop single-cell transcriptome solution.

Core Technology

Single Cell Full-length RNA Sequence Transcriptome-seq (scFAST-seq) allows full-length transcriptome analysis of thousands of fresh or fixed cells in a single experiment. In contrast to the current single-cell transcriptome assays based on oligo-dT reverse transcription, the method used in scFAST-seq provides coverage of the whole transcriptome. The sensitivity of the scFAST-seq technique assays depends primarily on the efficiency of reverse transcription, cDNA enrichment, and ribosome depletion.

First, cells and barcoded beads are added and react to generate emulsion droplets in the shaped channel of the SeekOne® Digital Droplet System. Reverse transcription is then performed by encapsulating the microbeads coupled to cell labels and semi-random primers with individual cells in a single droplet, resulting in cell-labelled cDNA fragments. Once the reverse transcription of each cDNA is complete, a template transition sequence (template switch oligo, TSO) is added to the 3' end of the cDNA using the end-transferase activity of the reverse transcriptase. The barcoded cDNA is then amplified in vitro, where the ribosomal RNA (rRNA) is depleted. Finally, a portion of the cDNA is fragmented, end-repaired, and ligated to a sequencing adaptor. The library is constructed and amplified using the unique dual-indexed polymerase chain reaction (PCR) primers for sequencing on Illumina or MGI platforms. In this way, the cDNA amplified by PCR would reflect both polyA and nonpolyA RNA sequences.

Product Features

Accurate single-cell transcriptome
Genotype to phenotype
Capturing non-coding RNA
Crossing the species barrier

Workflow

Step. 1
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1. Sample transport
Step. 2
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2. Tissue dissociation
Step. 3
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3. Single cell separation and labeling
Step. 4
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4. Library construction
Step. 5
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5. NGS sequencing
Step. 6
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6. Bioinformatic analysis
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Overview

Achieve full-length transcriptome detection without being limited by the 3' or 5' end.
Simultaneously obtain single-cell mutation and gene expression info, establishing the relationship btw. genotype and phenotype
In addition to mRNA, non-coding RNA without 3' poly A tails can also be captured.
Enable the detection of viruses and prokaryotes.
Allow the detection of isoform switching events of mRNA and IncRNA.
The inclusion of an rRNA blocker enables efficient depletion of rRNA.
Wide range of application in tumor research, organoid, lncRNA, RNA editing, cell treatment, virus study, development, etc.
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Content

1 × SeekOne® DD Chip S3 Kit for 8 tests
1 × SeekOne® DD scFAST-seq Barcoded Beads Kit for 8 tests
1 × SeekOne® DD scFAST-seq Reverse Transcription Kit for 8 tests
1 × SeekOne® DD Library Construction Kit for 8 tests
1 × SeekOne® DD scFAST-seq Cleanup Kit for 8 tests

Product name
SeekOne DD Single Cell Full-length RNA Sequence Transcriptome-seq Kit (scFAST-seq)
Manufacturer
SeekGene BioSciences Co., Ltd.
Catalog no.
K00801-08
Others
For Research Use Only. Not for use in diagnostic procedures.

Documents

Brochure-SeekOne_DD_scFAST-seq.pdf
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