Total Rna Isolation
Total Rna Isolation - Related Videos
17:25
RNA Extraction from Neuroprecursor Cells Using the Bio-Rad Total RNA Kit
0 Views •
Cited by 1 •
2007
09:43
Isolation and Characterization of RNA-Containing Exosomes
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Cited by 515 •
2012
This paper demonstrates methods for the isolation, purification and detection of exosomes, as well as techniques for analysis of their molecular content. These methods are adaptable for exosome isolation from both cell culture media and biological fluids, and can beyond analysis of molecular content also be useful in functional studies.
14:49
Single Read and Paired End mRNA-Seq Illumina Libraries from 10 Nanograms Total RNA
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Cited by 30 •
2011
Here we describe a method for preparation of both single read and paired end Illumina mRNA-Seq sequencing libraries for gene expression analysis based on T7 linear RNA amplification. This protocol requires only 10 nanograms of starting total RNA and generates highly consistent libraries representing whole transcripts.
10:06
Non-Laser Capture Microscopy Approach for the Microdissection of Discrete Mouse Brain Regions for Total RNA Isolation and Downstream Next-Generation Sequencing and Gene Expression Profiling
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Cited by 11 •
2011
RNA expression profiling of discrete mouse brain regions requires a precise and repeatable tissue collection strategy. A protocol that uses both coronal brain sectioning and tissue corer-assisted microdissection is described here. The yield and quality of total RNA obtained from the resulting samples confirms the utility of the outlined method.
11:09
Chromatin Isolation by RNA Purification (ChIRP)
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Cited by 380 •
2012
ChIRP is a novel and rapid technique to map genomic binding sites of long noncoding RNAs (lncRNAs). The method takes advantage of the specificity of anti-sense tiling oligonucleotides to allow the enumeration of lncRNA-bound genomic sites.
FAQs
The critical preservation step is rapid RNase inactivation after cells or tissues are disrupted. RNases can degrade RNA during handling, so effective inactivation helps maintain the transcript abundance present in the original sample. This matters because downstream measurements, including gene-expression studies and quantitative PCR, are only useful when the isolated RNA reflects the biological starting material.
These approaches separate RNA from unwanted cellular material through different physical and chemical principles. Chemical phase separation partitions components into distinct phases, silica-column binding captures RNA on a solid surface, and alcohol precipitation concentrates RNA from a solution. The selected approach determines how the sample is processed to remove DNA, proteins, lipids, and other contaminants.
Because the preparation retains messenger RNA, ribosomal RNA, and transfer RNA, it provides a broader view of the sample’s RNA composition than an approach focused on only one class. Messenger RNA supports gene-expression analysis, while the presence of other major classes contributes to the overall RNA preparation assessed for quality before downstream molecular applications.
A typical workflow begins by disrupting cells or tissues, followed by chemical conditions that inactivate RNases. The sample is then processed to separate RNA from DNA, proteins, lipids, and other cellular components using phase separation, silica binding, or alcohol precipitation. Finally, the recovered RNA is evaluated for quantity and integrity before molecular analysis.
Quantity indicates how much RNA the preparation contains, whereas integrity indicates whether the molecules remain sufficiently preserved for subsequent analysis. Considering both measurements helps researchers judge whether the sample is suitable for reverse transcription, quantitative PCR, RNA sequencing, or gene-expression studies. These checks connect the extraction outcome with the reliability of later molecular results.
Biologists use the resulting RNA preparations to investigate cellular regulation, development, disease mechanisms, and responses to experimental conditions. The material can support reverse transcription, quantitative PCR, RNA sequencing, and broader gene-expression studies. Its value lies in preserving RNA information from the biological sample so researchers can compare molecular states or examine changes associated with specific conditions.