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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


Service Details

Bioinformatics

Demo Results

Advantages

Ø Highly Experienced: Over 200,000 samples have been processed in BMK covering diverse sample types, including cell culture, tissue, body fluid, etc. and over 7,000 mRNA-Seq projects closed covering various research area.

Ø Strict quality control system: Core quality control points through all steps including sample preparation, library preparation, sequencing and bioinformatics are under close monitoring in order to deliver high-quality results.

Ø Multiple databases available for function annotation and enrichment studies to fulfill diverse research goals.

Ø After-sale services: After-sale services valid for 3 months upon project completion, including projects follow-up, trouble-shooting, results Q&A, etc.

Sample Requirements and Delivery

Library Sequencing strategy Data recommended Quality Control
Poly A enriched Illumina PE150

6 Gb

Q30≥85%

Sample Requirements:

Nucleotides:

Purity Integrity Amount
OD260/280≥1.7-2.5 OD260/230≥0.5-2.5Limited or no protein or DNA contamination shown on gel. For plants: RIN≥6.5;For animals: RIN≥7;28S/18S≥1.0;limited or no baseline elevation Conc. ≥30 ng/μl; Volumn ≥ 10 μl; Total ≥ 1.5 μg

Tissue: Weight(dry): ≥1 g
*For tissue smaller than 5 mg, we recommend to send flash frozen(in liquid nitrogen) tissue sample.

Cell suspension: Cell count = 3×106 - 1×107
*We recommend to ship frozen cell lysate. In case that cell counts smaller than 5×105, flash frozen in liquid nitrogen is recommended, which is preferable for micro extraction.

Blood samples: Volume≥1 ml

Microorganism: Mass ≥ 1 g

Recommended Sample Delivery

Container: 2 ml centrifuge tube (Tin foil is not recommended)

Sample labeling: Group+replicate e.g. A1, A2, A3; B1, B2, B3... ...

Shipment:

  1. Dry-ice: Samples need to be packed in bags and buried in dry-ice.
  2. RNAstable tubes: RNA samples can be dried in RNA stabilization tube(e.g. RNAstable®) and shipped in room temperature.

Service Work Flow

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Experiment design

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Sample delivery

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RNA extraction

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Library construction

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Sequencing

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Data analysis

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After-sale services


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  • Bioinformatics

    2116 (1)

    Eukaryotic mRNA sequencing analysis workflow

    Bioinformatics

    Ø  Raw data quality control

    Ø  Reference genome alignment

    Ø  Transcript structure analysis

    Ø  Expression quantification

    Ø  Diffferential expression analysis

    Ø  Function annotation and enrichment

    1.mRNA Data Saturation curve

    3(1)

    2.Differential expression analysis-Volcano plot

    4(1)

    3.KEGG annotation on DEGs

    5(1)

    4.GO classification on DEGs

    6(1)

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