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Small RNA sequencing-Illumina

Small RNA (sRNA) molecules, typically under 200 nucleotides in length, include microRNAs (miRNAs), small interfering RNAs (siRNAs), and piwi-interacting RNAs (piRNAs). Among these, miRNAs, around 20-24 nucleotides long, are particularly noteworthy for their pivotal regulatory roles in various cellular processes. With tissue-specific and stage-specific expression patterns, miRNAs exhibit high conservation across different species.

Platform: Illumina NovaSeq


Service Details

Bioinformatics

Demo Results

Featured Publications

Features

● Size selection of RNA prior to library preparation

● Bioinformatic analysis centered around miRNA prediction and their targets

Service Advantages

● Comprehensive bioinformatics analysis: enabling the identification of both known and novel miRNAs, identification of miRNAs targets and corresponding functional annotation and enrichment with multiple databases (KEGG, GO)

● 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.

● 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.

● Extensive Expertise: with a track record of successfully closing over multiple sRNA projects covering over 100 species in various research domains, our team brings a wealth of experience to every project.

Sample Requirements and Delivery

Library

Platform

Recommended data

Data QC

Size selected

Illumina SE50

10M-20M reads

Q30≥85%

Sample Requirements:

Nucleotides:

Conc.(ng/μl)

Amount (μg)

Purity

Integrity

≥ 80

≥ 0.5

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: 450 mg

   Viscera or Brain: 240 mg

   Muscle: 600 mg

   Bones, Hair or Skin: 1.5g

● Arthropods:

   Insects: 9g

   Crustacea: 450 mg

● Whole blood: 2 tubes

● Cells: 106 cells

● Serum and Plasma: 6 mL

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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    Identification of miRNA: structure and depth

     

     

     miRNA-precursor-structure-and-sequencing-depth

     

    Differential expression of miRNA – hiearchical clustering

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    Functional annotation of target of differentially expressed miRNAs

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    Explore the research advancements facilitated by BMKGene’ sRNA sequencing services through a curated collection of publications.

      

    Chen, H. et al. (2023) ‘Viral infections inhibit saponin biosynthesis and photosynthesis in Panax notoginseng’, Plant Physiology and Biochemistry, 203, p. 108038. doi: 10.1016/J.PLAPHY.2023.108038.

    Li, H. et al. (2023) ‘ The plant FYVE domain‐containing protein FREE1 associates with microprocessor components to repress miRNA biogenesis ’, EMBO reports, 24(1). doi: 10.15252/EMBR.202255037/SUPPL_FILE/EMBR202255037-SUP-0004-SDATAFIG4.TIF.

    Yu, J. et al. (2023) ‘The MicroRNA Ame-Bantam-3p Controls Larval Pupal Development by Targeting the Multiple Epidermal Growth Factor-like Domains 8 Gene (megf8) in the Honeybee, Apis mellifera’, International Journal of Molecular Sciences, 24(6), p. 5726. doi: 10.3390/IJMS24065726/S1.

    Zhang, M. et al. (2018) ‘Integrated Analysis of MiRNA and Genes Associated with Meat Quality Reveals that Gga-MiR-140-5p Affects Intramuscular Fat Deposition in Chickens’, Cellular Physiology and Biochemistry, 46(6), pp. 2421–2433. doi: 10.1159/000489649.

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