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Related Concept Videos

MicroRNAs01:22

MicroRNAs

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MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After...
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MicroRNAs01:22

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MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...
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mirMachine: A One-Stop Shop for Plant miRNA Annotation
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A method for validating microRNAs in plants by miR-RACE.

Jinggui Fang1, Xin Sun

  • 1College of Horticulture, Nanjing Agricultural University, Nanjing, Jiangsu Province, 210095, People's Republic of China, fanggg@njau.edu.cn.

Methods in Molecular Biology (Clifton, N.J.)
|March 6, 2015
PubMed
Summary

Accurate microRNA (miRNA) sequencing is crucial. The miRNA rapid amplification of cDNA ends (miR-RACE) method precisely maps mature miRNA sequences, overcoming limitations of prediction algorithms.

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Area of Science:

  • Molecular Biology
  • Genomics
  • Bioinformatics

Background:

  • Computational miRNA prediction algorithms often lack nucleotide-level precision in identifying mature miRNA locations within precursor sequences.
  • Accurate determination of mature miRNA sequences is essential for understanding their regulatory functions.

Purpose of the Study:

  • To present the miRNA rapid amplification of cDNA ends (miR-RACE) as an efficient method for precise miRNA sequence determination.
  • To provide a protocol for validating computationally predicted miRNA sequences.

Main Methods:

  • The miR-RACE protocol involves miRNA-enriched library preparation.
  • It utilizes two specific 5'- and 3'-miRNA RACE (miR-RACE) PCR reactions.
  • Sequence-directed cloning follows the PCR reactions.

Main Results:

  • The miR-RACE method enables precise sequencing of mature microRNAs.
  • The protocol is rapid, completable within 2-3 days.
  • Primer design for the RACE reactions is identified as a critical step.

Conclusions:

  • miR-RACE is an effective technique for validating and precisely locating computationally predicted miRNAs.
  • This method enhances the accuracy of miRNA identification in molecular biology research.
  • The rapid nature of miR-RACE facilitates timely experimental validation.