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Updated: Jul 5, 2026
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Tissue Collection of Bats for -Omics Analyses and Primary Cell Culture
Published on: October 23, 2019
Preparation of RNA from tissues and cells
R Ribaudo1, M Gilman, R E Kingston
1National Institute of Allergy and Infectious Disease, Bethesda, Maryland, USA.
Current Protocols in Neuroscience
|April 23, 2008
Summary
This study presents methods for isolating RNA from eukaryotic cells, including hot phenol extraction and rapid cytoplasmic RNA preparation. These techniques minimize DNA contamination for accurate molecular biology research.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Isolating RNA from eukaryotic cells is crucial for molecular studies.
- Standard RNA isolation often requires DNA removal steps.
- Contamination by DNA can interfere with downstream RNA analysis.
Purpose of the Study:
- To describe efficient methods for RNA isolation from eukaryotic cells.
- To minimize DNA contamination during the RNA extraction process.
- To provide protocols for total RNA, cytoplasmic RNA, and mRNA isolation.
Main Methods:
- Hot phenol extraction to shear and remove DNA.
- Nonionic detergent lysis for cytoplasmic RNA, followed by centrifugation to remove nuclei.
- Guanidinium thiocyanate and CsCl step gradient for separating RNA from other macromolecules.
- Protocol for isolating poly(A(+)) messenger RNAs (mRNAs) from total RNA.
Main Results:
- Hot phenol extraction effectively reduces DNA contamination.
- Rapid cytoplasmic RNA preparation isolates RNA while leaving nuclei intact.
- Guanidinium thiocyanate protocol achieves high-purity RNA separation.
- Specific protocol enables isolation of poly(A(+)) mRNAs.
Conclusions:
- Multiple effective protocols exist for eukaryotic RNA isolation.
- These methods address DNA contamination challenges.
- The described techniques facilitate various RNA research applications, including mRNA isolation.
