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RNA-seq

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...

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Method for the Isolation and Identification of mRNAs, microRNAs and Protein Components of Ribonucleoprotein Complexes from Cell Extracts using RIP-Chip

Method for the Isolation and Identification of mRNAs, microRNAs and Protein Components of Ribonucleoprotein Complexes from Cell Extracts using RIP-Chip

Published on: September 29, 2012

Microchip-based solid-phase purification of RNA from biological samples.

Kristin A Hagan¹, Joan M Bienvenue, Christopher A Moskaluk

¹Department of Chemistry, University of Virginia, Charlottesville, Virginia 22904, USA.

Analytical Chemistry

|October 16, 2008

Summary

This summary is machine-generated.

This study presents a rapid, closed-system microchip method for purifying RNA from biological samples using silica beads. The efficient RNA extraction method is suitable for subsequent amplification, including reverse transcription PCR (RT-PCR).

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Method for the Isolation and Identification of mRNAs, microRNAs and Protein Components of Ribonucleoprotein Complexes from Cell Extracts using RIP-Chip

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Published on: January 3, 2019

Area of Science:

Biotechnology
Analytical Chemistry
Molecular Biology

Background:

Accurate RNA purification is crucial for sensitive molecular analyses.
Existing RNA extraction methods can be time-consuming and prone to contamination.
A need exists for rapid, efficient, and closed-system RNA purification techniques.

Purpose of the Study:

To demonstrate a microchip-based solid-phase extraction method for RNA purification.
To determine the binding capacity of the microchip device for RNA.
To evaluate the efficiency and suitability of the purified RNA for downstream applications like RT-PCR.

Main Methods:

Development of a microchip device utilizing silica beads for solid-phase extraction of RNA.
Determination of the device's binding capacity for RNA, RNA with protein, and DNA.
Assessment of RNA purification time and reagent consumption compared to commercial methods.
Validation of purified RNA integrity and amplifiability using reverse transcription PCR (RT-PCR).

Main Results:

The microchip device demonstrated a binding capacity of 360 ng for RNA in the presence of protein.
RNA extraction was achieved in approximately 9 minutes, with reduced reagent consumption.
The closed-system design minimized the risk of RNase and contaminant introduction.
Purified RNA was successfully amplified via RT-PCR, confirming its suitability for genetic analysis.

Conclusions:

The microchip-based RNA purification method offers a rapid, efficient, and closed-system alternative to commercial methods.
The device exhibits sufficient capacity and yields amplifiable RNA, making it valuable for genetic analysis of various biological samples.
This technology has potential applications in forensic science and clinical diagnostics, particularly for sensitive RNA-based analyses.