Eukaryotic mRNA Sequencing-Illumina
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 closed 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 the corresponding enrichment analysis, providing insights on 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:
- Dry-ice: Samples need to be packed in bags and buried in dry-ice.
- RNAstable tubes: RNA samples can be dried in RNA stabilization tube(e.g. RNAstable®) and shipped in room temperature.
Service Work Flow
Experiment design
Sample delivery
RNA extraction
Library construction
Sequencing
Data analysis
After-sale services
Bioinformatics
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
Data Saturation
Sample correlation and assessment of biological replicates
Differentially Expressed Genes (DEGs)
Functional Annotation of DEGs
Functional Enrichment of DEGs
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.
