Whole Transcriptome Amplification
Whole Transcriptome Amplification - Related Videos
09:44
Rapid Amplification of cDNA Ends
0 Views •
2023
Source: Pablo Sanchez Bosch2, Sean Corcoran2 and Katja Brückner1,2,3 1Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research 2Department of Cell and Tissue Biology, 3Cardiovascular Research Institute, University of California San Francisco, San Francisco, CA, USA Rapid Amplification of cDNA Ends (RACE) is a technique that allows amplification of full-length cDNA from mRNA by extending to the 3’ or 5’ end, even without prior knowledge of the sequence (Frohman et al.,...
07:27
Transcriptome Analysis of Single Cells
0 Views •
Cited by 65 •
2011
In this article we describe a simple method for the harvesting of single cells from rat primary neuronal cultures and subsequent transcriptome analysis using aRNA amplification. This approach is generalizable to any cell type.
10:12
Protein Misfolding Cyclic Amplification of Prions
0 Views •
Cited by 7 •
2012
Protein misfolding cyclic amplification (PMCA) is an in vitro assay for the study of prion conversion and strain and species barriers. It can also be used as a prion detection assay.
02:41
Amplification of a Suicide Plasmid in a Bacterial Host
0 Views •
2025
Source: Tomás, J. M., et al. Generation of Null Mutants to Elucidate the Role of Bacterial Glycosyltransferases in Bacterial Motility. J. Vis. Exp. (2022).This video demonstrates the initiator-dependent amplification of a suicide plasmid in E. coli. The plasmid, which carries an antibiotic resistance gene, replicates only in the presence of an initiator protein encoded by the host. After electroporation and incubation in recovery medium, the mixture is plated on antibiotic-containing agar.
13:51
Analyzing Gene Expression from Marine Microbial Communities using Environmental Transcriptomics
0 Views •
Cited by 60 •
2009
We present a method for generating cDNA from environmental mRNA. In general, total RNA is first collected from the environment, rRNA is selectively removed, mRNA is selectively amplified, and cDNA synthesized from the enriched mRNA pool is sequenced. Recovered sequences can be annotated using standard bioinformatics techniques to identify the expressed genes.
FAQs
The workflow first converts RNA into complementary DNA, or cDNA, creating a representation of the original transcripts that can undergo further synthesis. Primer-directed copying then generates product across many transcript sequences rather than focusing on one gene. This expanded, transcriptome-wide material provides enough input for downstream sequencing or other assays when the starting RNA is limited.
Linear and isothermal amplification provide alternative routes for generating sufficient transcriptome-wide product after cDNA formation and primer-directed synthesis. Both strategies support the central goal of increasing RNA-derived material across many transcripts. Their distinction lies in how amplification proceeds, while the intended outcome remains a broad product suitable for subsequent transcriptome analysis.
Maintaining relative expression patterns helps the amplified product remain informative about differences among transcripts in the original sample. That feature is important for gene-expression profiling because researchers need to interpret which transcripts are represented more or less strongly. Preserving these relationships makes the expanded material more useful for characterizing cellular states and molecular responses.
A typical workflow begins with RNA from a biological sample and converts it into complementary DNA. Primer-directed synthesis then amplifies material representing many transcripts, using either a linear or an isothermal strategy. The resulting transcriptome-wide product can be directed into sequencing or another assay, extending the analysis beyond what the original RNA quantity would permit.
WTA is particularly useful when a sample provides too little RNA for direct transcriptome analysis. This includes scarce material from single cells, rare specimens, or archived samples. By increasing the available RNA-derived product, the method allows researchers to examine transcript information that might otherwise be inaccessible and supports broader characterization of the sample.
The product can support gene-expression profiling by making transcriptome-wide material available for sequencing or other assays. Analyses may reveal patterns associated with cellular states or molecular responses, rather than limiting the investigation to a single transcript. The value of the output therefore depends on how well it represents the many transcripts present in the starting sample.
In biology, WTA extends transcriptome analysis to settings where material is scarce, including single-cell studies, developmental biology, disease research, and investigations of rare or archived specimens. These applications use expanded transcript information to characterize changing cellular states and molecular responses. The method therefore connects limited biological samples with broader gene-expression measurements.