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Eukaryotic mRNA Sequencing-Illumina

mRNA sequencing empowers the comprehensive profiling of all mRNA transcripts within cells under specific conditions. This cutting-edge technology serves as a potent tool, unveiling intricate gene expression profiles, gene structures, and molecular mechanisms associated with diverse biological processes. Widely adopted in fundamental research, clinical diagnostics, and drug development, mRNA sequencing offers insights into the intricacies of cellular dynamics and genetic regulation.

Platform: Illumina NovaSeq X


Service Details

Bioinformatics

Demo Results

Featured Publications

Features

● Capture of poly mRNA prior to library preparation

● Optional directional mRNA library preparation to enable obtaining strand-specific sequencing data

● Bioinformatic analysis of gene expression and transcript structure

 

Advantages

● Extensive Expertise: with a track record of processing over 200,000 samples at BMK, spanning diverse sample types such as cell cultures, tissues, and body fluids, our team brings a wealth of experience to every project. We've successfully completed over 10,000 mRNA-Seq projects in various research domains.

● Rigorous Quality Control: we implement core control points across all stages, from sample and library preparation to sequencing and bioinformatics. This meticulous monitoring ensures the delivery of consistently high-quality results.

● Comprehensive Annotation: we use multiple databases to functionally annotate the Differentially Expressed Genes (DEGs) and perform corresponding enrichment analysis, providing insights into the cellular and molecular processes underlying the transcriptome response.

● Post-Sales Support: Our commitment extends beyond project completion with a 3-month after-sale service period. During this time, we offer project follow-up, troubleshooting assistance, and Q&A sessions to address any queries related to the results.

Sample Requirements and Delivery

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

6 -10 Gb

Q30≥85%

Sample Requirements:

Nucleotides:

 

Conc.(ng/μl)

Amount (μg)

Purity

Integrity

Standard Library

≥ 20

≥ 0.5

OD260/280=1.7-2.5

OD260/230=0.5-2.5

Limited or no protein or DNA contamination shown on gel.

For plants: RIN≥6.0;

For animals: RIN≥6.5;

 5.0≥28S/18S≥1.0;

limited or no baseline elevation

Directional Library

≥ 30

≥ 1

OD260/280=1.7-2.5

OD260/230=0.5-2.5

Limited or no protein or DNA contamination shown on gel.

RIN≥6.5;

 5.0≥28S/18S≥1.0;

limited or no baseline elevation

● Plants:

   Root, Stem or Petal: 450 mg

   Leaf or Seed: 300 mg

   Fruit: 1.2 g

  Animal:

   HEart or Intestine: 300 mg

   Viscera or Brain: 240 mg

   Muscle: 450 mg

   Bones, Hair or Skin: 1g

●  Arthropods:

   Insects: 6g

   Crustacea: 300 mg

● Whole blood: 1 tube

●  Cells: 106 cells

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

Sample QC

Experiment design

sample delivery

Sample delivery

Pilot experiment

RNA extraction

Library Preparation

Library construction

Sequencing

Sequencing

Data analysis

Data analysis

After sale Services

After-sale services


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  • Bioinformatics

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    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

      Reference Genome Alignment

    图片1

     

    Data Saturation 

    3(1)

     

     

    Sample correlation and assessment of biological replicates

    图片2

     

    Differentially Expressed Genes (DEGs)

     

    4(1)

     

     

    Functional Annotation of DEGs

     

    6(1)

     

     

     

    Functional Enrichment of DEGs

    图片4

     

    Explore the advancements facilitated by BMKGene’s eukaryotic NGS mRNA sequencing services through a curated collection of publications.

     

    Huang, L. et al. (2023) ‘Triclosan and triclocarban weaken the olfactory capacity of goldfish by constraining odorant recognition, disrupting olfactory signal transduction, and disturbing olfactory information processing’, Water Research, 233, p. 119736. doi: 10.1016/J.WATRES.2023.119736.

    Jia, L. J. et al. (2023) ‘Aspergillus fumigatus hijacks human p11 to redirect fungal-containing phagosomes to non-degradative pathway’, Cell Host & Microbe, 31(3), pp. 373-388.e10. doi: 10.1016/J.CHOM.2023.02.002.

    Jin, K. et al. (2022) ‘TCP Transcription Factors Involved in Shoot Development of Ma Bamboo (Dendrocalamus latiflorus Munro)’, Frontiers in Plant Science, 13, p. 884443. doi: 10.3389/FPLS.2022.884443/BIBTEX.

    Wen, X. et al. (2022) ‘The Chrysanthemum lavandulifolium genome and the molecular mechanism underlying diverse capitulum types’, Horticulture Research, 9. doi: 10.1093/HR/UHAB022.

    Zhang, Yujie et al. (2023) ‘A cascade nanoreactor for enhancing sonodynamic therapy on colorectal cancer via synergistic ROS augment and autophagy blockage’, Nano Today, 49, p. 101798. doi: 10.1016/J.NANTOD.2023.101798.

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