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Eukaryotic Transcriptome Sequencing with Reference Genome

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Introductions

Eukaryote Transcriptome Sequencing with Reference Genome

Transcriptome refers to the collection of all transcripts of a specific cell or tissue of a species in a certain state. A transcriptome with a reference genome refers to a sequencing project where the reference genome of a species has been published, and we can use the known reference for analysis. Transcriptome sequencing with reference genomes can not only quantitatively determine the changes in the expression level of each transcript during growth and under different conditions, but also analyze the gene structure and study SNPs and alternatively splicing, gene structure optimization and other information. Through the new generation of high-throughput sequencing, almost all transcript sequence information of a specific tissue or organ of a species in a certain state can be obtained comprehensively and quickly, and it has been widely used in agriculture, medical research, food and other fields.

Services

With years of rich experience, Lifeasible provides transcriptome sequencing service with reference genome. We provide a one-stop service model. We complete a series of processes from sample extraction, quality testing, sequencing, biological information analysis to making reports.

Workflow of transcriptome sequencing with reference genome

Workflow of transcriptome sequencing with reference genome

Analysis content

Basic analysis

  • Sequencing quality evaluation and raw data filtering, removing adapter sequences and low-quality reads
  • Align reference genome or reference gene sequence
  • Sequencing and comparison evaluation (data comparison statistics, sequencing saturation analysis, sequencing randomness analysis)
  • Gene expression statistics (gene coverage, expression, expression abundance distribution)
  • Prediction and annotation of novel gene transcripts (reference genome is required, limited to animals and plants)
  • Sample relationship analysis (principal component analysis, correlation test, sample cluster diagram)
  • Differentially expressed genes analysis (two or more samples)

Screening of differentially expressed genes

Volcano map and cluster analysis of expression patterns (heat map) of differential genes

Significant enrichment analysis of GO/KEGG function of differential genes

Reactome/DO significant enrichment analysis of differential genes (limited to some species)

GSEA analysis (GO/KEGG/Reactome/DO)

Advanced analysis

  • Gene structure optimization
  • Gene alternative splicing identification
  • Trend analysis

Sample requirements

  • Total sample requirement≥5μg, concentration≥100ng/μl
  • Animal sample purity RIN≥8, OD260/280≥1.8, 28S/18S≥1.0
  • Please put the sample in a 1.5ml tube, indicate the sample name, concentration and preparation time on the tube, and use parafilm to seal the tube mouth.
  • Before shipping, fix all sample tubes in 50ml centrifuge tubes with caps and ship them on dry ice.

Project cycle

  • The entire project generally takes about 2 months.

Feedback to customers

  • Raw data
  • A project report includes experimental process, instrument parameters, analysis methods and results, etc.

Service process

Service process

Advantages

  • From material selection, library construction and sequencing, to data analysis, every step has a scientific and meticulous design to ensure high-quality research results.
  • The one-stop service we provide makes your upstream and downstream experiments seamlessly connected, facilitates your experiments more conveniently and quickly.
  • The continuously updated and expanded high-performance computing platform ensures efficient data processing and secure data storage.
  • Personalized customized package service.
  • Standardization system, scientific management.

※ It should be noted that our service is only used for research.

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