Microrna Sequencing
Microrna Sequencing - Related Videos
01:22
MicroRNAs
0 Views •
2019
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 ends...
09:19
In ovo Expression of MicroRNA in Ventral Chick Midbrain
0 Views •
Cited by 5 •
2013
Ectopic expression is one technique to elucidate the microRNAs role in brain development. However, targeting specific areas using in ovo electroporation is challenging. Here, we show an efficient way to selectively electroporate ventral and dorsal midbrain regions.
00:09
A Complete Pipeline for Isolating and Sequencing MicroRNAs, and Analyzing Them Using Open Source Tools
0 Views •
Cited by 3 •
2019
Here, we describe a step-by-step strategy for isolating small RNAs, enriching for microRNAs, and preparing samples for high-throughput sequencing. We then describe how to process sequence reads and align them to microRNAs, using open source tools.
10:55
Isolating, Sequencing and Analyzing Extracellular MicroRNAs from Human Mesenchymal Stem Cells
0 Views •
Cited by 3 •
2019
This protocol demonstrates how to purify extracellular microRNAs from cell culture media for small RNA library construction and next generation sequencing. Various quality control checkpoints are described to allow readers to understand what to expect when working with low input samples like exRNAs.
07:23
Describing a Transcription Factor Dependent Regulation of the MicroRNA Transcriptome
0 Views •
Cited by 3 •
2016
Herein we propose a strategy to study the effect of a transcription factor of interest on the microRNA transcriptome using publically available data, computational resources and high throughput data from microRNA arrays after transfecting cells with small hairpin (sh)RNA targeting a transcription factor of interest.
FAQs
These steps convert mature microRNAs into a sequenceable library. Adapter ligation adds the sequences needed to process the small RNA molecules, reverse transcription converts the RNA-based information into a form suitable for library processing, and amplification increases the available library material before sequencing. Together, they connect the original molecules to measurable sequence data.
After high-throughput sequencing, computational alignment assigns resulting reads to reference genomes or microRNA databases. This connects individual sequence reads with recognized microRNA sequences and supports their quantification across samples. The organized profile then allows researchers to compare microRNA abundance between biological conditions rather than examining raw sequence information alone.
MicroRNAs regulate gene expression after transcription, so altered abundance can provide clues about changes in cellular programs. In cancer research, sequencing profiles may highlight microRNAs associated with proliferation, invasion, metastasis, or treatment response. These associations help researchers investigate regulatory pathways and prioritize molecular patterns for further study.
A typical workflow begins with mature microRNA molecules and builds sequencing libraries through adapter ligation, reverse transcription, and amplification. The libraries then undergo high-throughput sequencing, after which computational analysis aligns the reads to reference genomes or databases and quantifies the detected microRNAs. The final output is a comparative expression profile.
This comparison is useful when investigators want to identify tumor-associated microRNA signatures. Differences between normal and malignant tissue profiles can reveal patterns linked to transformation or tumor behavior, while the broader expression contrast helps distinguish cancer-associated regulation from microRNA activity present in nonmalignant tissue. Such findings can support biomarker discovery.
The resulting profiles can support discovery of candidate biomarkers and improve understanding of post-transcriptional regulation in cancer. Researchers can also examine whether microRNA patterns differ with tumor-associated processes or treatment response. These outcomes do not replace biological validation, but they provide a detailed molecular basis for selecting signatures and pathways for additional investigation.