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Small RNA Sequencing Service

Importance Workflow Samples Bioinformatics Analysis Advantages Q&As Resource

Equipped with world-leading technology platforms and professional scientific staff, Creative Biolabs is a world-leading services provider in the field of next-generation sequencing (NGS). Based on our versatile SuPrecision™ platform, we offer one-stop small RNA sequencing services for global customers from small RNA extraction to data analysis. We are committed to delivering accurate, affordable and high-quality sequencing data to support your research.

Importance of Small RNA Sequencing in Cancer Research

Small RNA species are a class of lowly abundant, non-coding RNA molecules with a length of less than 200 nt. The kinds of small RNA include microRNA (miRNA), small interfering RNA (siRNA), Piwi-interacting RNA (piRNA), small nucleolar RNA (snoRNAs), tRNA-derived small RNA (tsRNA), small rDNA-derived RNA (srRNA) and small nuclear RNA (snRNA). These small RNAs play essential roles in gene silencing and post-transcriptional regulation of gene expression, involved in a number of biological processes including cell proliferation and differentiation, and apoptosis. Currently, accumulated evidence demonstrated that small RNAs are associated with the development and progression of cancer, as well as anti-tumor drug resistance. For example, the over-expression of miR-17 was associated with chemo-resistant colon cancer. miR-34 was involved in the regulation of drug resistance in gastric, prostate and breast cancers. The upregulated levels of piR-4987, piR-20365, piR-20485 and piR-20582 were found in tumor tissue. Hence, to explore small RNAs’ roles in oncology will increase the understanding of tumor pathological progression and accelerate discovery of novel anti-tumor therapies.

Small RNA-Seq is a powerful method that can quickly identify the known small RNAs’ level under specific conditions and discover novel small RNA molecules. Meanwhile, it allows small RNAs’ profiling under different conditions and the correlation analysis with transcriptome sequencing expression profile data.

Small RNA-Seq Workflow at Creative Biolabs

Total RNA
Isolation
Small RNA
Enrichment
Small RNA Library Construction
Sequencing
Bioinformatics
Analysis

Sample Requirements

Tissue or cell Sample:

1) Fresh cultured cell (number of cells): ≥ 2×10⁵ cells
2) Fresh animal tissue (dry weight): ≥ 30 mg
3) Whole blood: ≥ 3 ml
4) FFPE: 5 pieces, 100 mm², 5~10 μm thick, unstained

RNA Sample:

1) Amounts of total RNA: ≥ 10 μg;
2) Concentration of total RNA: ≥ 200 ng/μL;
3) RNA purity: OD260/280=1.8~2.2, OD260/230 ≥ 2.0, RIN ≥ 6.5, 28S:18S ≥ 1.0;
4) Amounts of small RNA: ≥ 1 μg;
5) Concentration of small RNA: ≥ 20 ng/μL.

Note! There are differences in sample requirements between different samples. For details, please consult our scientists.

Bioinformatics Analysis

miRNA nucleotide bias at each position. (Creative Biolabs Authorized)

Standard Analyses

1) QC for raw data
Sequence alignment
2) Small RNA classification and annotation
3) Analysis for known miRNA (including expression levels, length distribution, and base preference)
4) Functional element annotation
5) Prediction for novel small RNA
6) Analysis of small RNA expression difference between samples
7) Target gene prediction
8) GO annotation and KEGG passway analysis for differentially expressed small RNA target gene
...

Senior Analyses

1) Prediction for miRNA target gene (non-model species or new miRNA)
2) Analysis of the differential expression of novel miRNA
3) Correlation analysis between differentially expressed miRNA and differentially expressed target genes
4) Interaction network between differentially expressed miRNA and differentially expressed target genes
5) Functional clustering analysis of interaction target genes (KEGG and GO)
...

Advantages

More than 3 million sequences can be obtained by one sequencing
Not only identify the known small RNA but also discover novel small RNA
Enable to detect the difference of single base of small RNA
High reliable and repeatable data

Creative Biolabs provides small RNA sequencing service packages including sample preparation, library construction, deep sequencing, raw data quality control, and customized bioinformatics analysis. If you have additional requirements or questions, please feel free to contact us.

Q&As

Q: How does small RNA sequencing help in understanding the role of microRNAs in cancer?

A: Small RNA sequencing identifies and quantifies microRNAs (miRNAs), which are crucial regulators of gene expression. By analyzing miRNA profiles, researchers can understand their roles in cancer development, progression, and metastasis, offering insights into potential therapeutic targets and biomarkers.

Q: Can small RNA sequencing detect novel small RNAs in cancer?

A: Yes, small RNA sequencing is capable of identifying novel small RNAs that have not been previously annotated. This is particularly useful in cancer research, where discovering new small RNAs can lead to a better understanding of cancer biology and the identification of new biomarkers and therapeutic targets.

Q: What is the importance of sequencing depth in small RNA sequencing?

A: Sequencing depth is crucial for accurately detecting and quantifying small RNAs, especially those with low expression levels. A higher sequencing depth ensures comprehensive coverage and reliable identification of small RNAs, which is essential for detailed cancer research.

Q: What are the advantages of using small RNA sequencing over other methods?

A: Small RNA sequencing offers higher sensitivity and specificity compared to traditional methods like microarrays. It allows for the discovery of novel small RNAs and provides a comprehensive view of the small RNA transcriptome, enabling a deeper understanding of gene regulation in cancer.

Q: Can small RNA sequencing be integrated with other omics data?

A: Yes, small RNA sequencing can be integrated with other omics data, such as genomics, transcriptomics, and proteomics. This integrative approach provides a comprehensive view of the molecular landscape in cancer, enhancing the understanding of disease mechanisms and aiding in the development of targeted therapies.