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Single- nucleus RNA Sequencing
The advance in single cell capturing and individual library construction technique combining with high-throughput sequencing allows gene expression studies on cell-by-cell basis. It enables a deeper and complete system analysis on complex cell populations, in which it largely avoids masking of their heterogeneity by taking average of all cells.
However, some cells are not suitable to be made into single-cell suspension, so other sample preparation methods – nucleus extraction from tissues are needed, that is, nucleus is directly extracted from tissues or cell and prepared into single-nucleus suspension for single-cell sequencing.
BMK provides 10× Genomics ChromiumTM based single-cell RNA sequencing service. This service has been widely used in studies on disease related studies, such as immune cell differentiation, tumor heterogeneity, tissue development, etc.
Spatial transcriptome chip: 10× Genomics
Platform: Illumina NovaSeq 6000
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BMKMANU S1000 Spatial Transcriptome
Spatial organization of cells plays vital role in various biological processes, such as immune infiltration, embryo development, etc. Spatial transcriptome sequencing, which indicates gene expression profiling while retaining information of spatial position, has provide great insights into transcriptome-level tissue architecture. With the development of technology, the ultra-clear tissue morphology and the real structural difference of spatial molecular expression need to be studied with higher resolution. BMKGENE provides comprehensive, one-stop spatial transcriptome sequencing service from samples to biological insights.
Spatial transcriptomics technologies empowered novel perspectives in diverse research arena by resolving gene expression profile with spatial content in heterogenous samples.
Spatial transcriptome chip: BMKMANU S1000
Platform: Illumina NovaSeq 6000
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10x Genomics Visium Spatial Transcriptome
Visium Spatial Gene Expression is a mainstream spatial transcriptome sequencing technology for classifying tissue based on total mRNA. Map the whole transcriptome with morphological context to discover novel insights into normal development, disease pathology, and clinical translational research. BMKGENE provides comprehensive, one-stop spatial transcriptome sequencing service from samples to biological insights.
Spatial transcriptomics technologies empowered novel perspectives in diverse research arena by resolving gene expression profile with spatial content in heterogenous samples.
Spatial transcriptome chip: 10x Genomics Visium
Platform: Illumina NovaSeq 6000
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Full-Length mRNA Sequencing-Nanopore
RNA sequencing has been an invaluable tool for comprehensive transcriptome analysis. Doubtlessly, traditional short-read sequencing achieved numerous important development in here. Nevertheless, it often encounters limitations in full-length isoform identifications, quantification, PCR bias.
Nanopore sequencing distinguishes itself from other sequencing platforms, in that the nucleotides are read directly without DNA synthesis and generates long read at tens of kilobases. This empowers direct read-out crossing full-length transcripts and tackling the challenges in isoform-level studies.
Platform： Nanopore PromethION
Library: cDNA-PCR
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Full-length mRNA sequencing -PacBio
De novo full-length transcriptome sequencing, also known as De novo Iso-Seq takes the advantages of PacBio sequencer in read length, which enables sequencing of full-length cDNA molecules without any breaks. This completely avoids any errors generated in transcript assembly steps and constructs unigene sets with isoform-level resolution. This unigene sets provides powerful genetic information as “reference genome” at transcriptome-level. In addition, combining with next generation sequencing data, this service empowers an accurate quantification of isoform-level expression.
Platform： PacBio Sequel IILibrary: SMRT bell library -
Eukaryotic mRNA Sequencing-Illumina
mRNA sequencing enables profiling of all mRNAs transcripted from cells under specific conditions. It is a powerful technology for revealing gene expression profile, gene structures and molecular mechanisms of certain biological processes. To date, mRNA sequencing has been widely employed in fundamental research, clinical diagnostics, drug development, etc.
Platform: Illumina NovaSeq 6000
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Non-Reference based mRNA Sequencing-Illumina
mRNA sequencing adopts next-generation sequencing technique (NGS) to capture the messenger RNA(mRNA) form Eukaryote at specific period that some special functions are activating. The longest transcript spliced was called ‘Unigene’ and used as the reference sequence for subsequent analysis, which is an effective means to study the molecular mechanism and regulatory network of the species without reference.
After transcriptome data assembly and unigene functional annotation
(1)SSR analysis, CDS prediction and gene structure will be preformed.
(2)Quantification of unigene expression in each sample will be performed.
(3)Differentially expressed unigenes between samples (or groups) will be discovered based on unigene expression
(4)Clustering, functional annotation and enrichment analysis of differentially expressed unigenes will be performed
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Long non-coding sequencing-Illumina
Long non-coding RNAs (lncRNAs) are a type of RNA molecules with length exceeding 200 nt, which are characterize by extremely low coding potential. LncRNA, as a key member in non-coding RNAs, is mainly found in nucleus and plasma. The development in sequencing technology and bioinformtics enables identification of numerous novel lncRNAs and associate those with biological functions. Accumulative evidences suggest that lncRNA is widely involved in epigenetic regulation, transcription regulation and post-transcription regulation.
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Small RNA sequencing-Illumina
Small RNA refers to a class of non-coding RNA molecules that are usually less than 200nt in length, including micro RNA (miRNA), small interference RNA (siRNA), and piwi-interacting RNA (piRNA).
MicroRNA (miRNA) is a class of endogenous small RNA with a length of about 20-24nt, which plays a variety of important regulatory roles in cells. miRNA involved in many life processes which revealing tissue – specific and stage – specific expression and highly conserved in different species.
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circRNA sequencing-Illumina
Whole transcriptome sequencing is designed to profile all types of RNA molecules, including coding (mRNA) and non-coding RNAs (including lncRNA, circRNA and miRNA) which are transcribed by specific cells at a certain time. Whole transcriptome sequencing, also known as “total RNA sequencing” aims at revealing comprehensive regulatory networks at transcriptome level. Taking advantage of NGS technology, sequences of entire transcriptome products are available for ceRNA analysis and joint RNA analysis, which provides the first step towards functional characterization. Revealing regulatory network of circRNA-miRNA-mRNA based ceRNA.
