miRNA Pull-down Service

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miRNA Pull-down Service

IntegrateRNA, a leading global biotechnology company specializing in RNA research, recently introduced the latest optimized miRNA pull-down strategy to help you identify the direct interactions between miRNAs and their target mRNAs in a faster and more accurate way. Our experienced team can provide flexible solutions for any individual request.

MicroRNAs (miRNA) are small non-coding regulatory RNAs, controlling key biological processes in a broad range of eukaryotic cells. The predominant function of miRNAs is to act as guide molecules in the RNA-induced silencing complexes (RISC) and target regions of complementarity in the mRNAs. Although bioinformatic tools have been employed to predict sequences within the 3' UTR of mRNAs as potential targets for miRNA binding. These computational methods often generate false positive results and are limited to predicting canonical interaction between miRNA and mRNA. Therefore, experimental procedures that measure direct binding of miRNA to its mRNA target are necessary to establish functional interaction. miRNA pull-down is a powerful tool to identify miRNA targets.

Strategy for Biotinylated miRNA Pull-down Assay

miRNA Pull-down Assay

As shown, we use biotinylated miRNA mimics as bait to enrich miRNA-mRNA complex, and streptavidin-coated magnetic beads absorb and screen the complex, which is more conducive to direct capture than antibody enrichment.

Workflow of miRNA Pull-down Service

  • Preparation of 3' biotinylated miRNA mimic (designed and synthesis based on the miRNA of your interest)
  • Transfection of biotinylated miRNA and control miRNA
  • Harvesting whole-cell lysate 48 hours after transfection
  • Incubating with streptavidin beads
  • Elution and separation of the pull-down products (containing miRNAs, mRNAs, RBPs)
  • Total RNA extraction and target detection (qRT-PCR, microarray and next-generation sequencing)
  • Data analysis and graph production

Advantages

  • Capture mRNA-miRNA complexes directly with end-labeled RNA, does not require antibodies for pull-down
  • High specificity and low bead background
  • Cost-effective one-stop solution
  • Raw data (including input, positive and negative control results), complete analytical data, SCI standard images

IntegrateRNA has many years of experiments in RNAs research and is dedicated to providing a comprehensive range of various types of RNA products and services for academic research, biotechnology research, and drug discovery. Based on the powerful platform and advance technique, we provide miRNA pull-down assay service to help you reveal the interaction mechanism between mRNA and miRNA. We guarantee our clients the most reliable and efficient research services to best match your research goals with faster turnaround time, lower prices and more comprehensive data analysis. If you have any questions, please feel free to contact us.

Reference:

  1. Sabyasachi D. et al. Biotin-based Pulldown Assay to Validate mRNA Targets of Cellular miRNAs. Journal of Visualized Experiments, 2018(136).
For research use only. Not intended for any clinical use.
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