ArticleNucleic acids research1996
Screening of differentially amplified cDNA products from RNA arbitrarily primed PCR fingerprints using single strand conformation polymorphism (SSCP) gels.
Article in Nucleic acids research, 1996. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 74 citations in OpenAlex.
- Trypanosomatid comparative genomics: Contributions to the study of parasite biology and different parasitic diseases.Genetics and molecular biology · 2012Article
- Cell culture and passaging alters gene expression pattern and proliferation rate in rheumatoid arthritis synovial fibroblasts.Arthritis research & therapy · 2010Article
- Cloning and identification of differentially expressed transcripts in primary culture of GABAergic neurons.Neurochemical research · 2001Article
- Applications of differential-display reverse transcription-PCR to molecular pathogenesis and medical mycology.Clinical microbiology reviews · 2000Review
- Prevention of depurination during elution facilitates the reamplification of DNA from differential display gels.Nucleic acids research · 1999Article
- Microglial activation varies in different models of Creutzfeldt-Jakob disease.Journal of virology · 1999Article
- Kinesin-like protein CENP-E is upregulated in rheumatoid synovial fibroblasts.Arthritis research · 1999Article
- Reciprocal subtraction differential RNA display: an efficient and rapid procedure for isolating differentially expressed gene sequences.Proceedings of the National Academy of Sciences of the United States of America · 1998Article
- Differential gene expression in response to adjunctive recombinant human interleukin-2 immunotherapy in multidrug-resistant tuberculosis patients.Infection and immunity · 1998Article
- Multiple isoform recovery (MIR)-PCR: a simple method for the isolation of related mRNA isoforms.Nucleic acids research · 1998Article
- Rapid determination of the complexity of cDNA bands extracted from DDRT-PCR polyacrylamide gels.Nucleic acids research · 1997Article
- Analysis of ammonia-oxidizing bacteria of the beta subdivision of the class Proteobacteria in coastal sand dunes by denaturing gradient gel electrophoresis and sequencing of PCR-amplified 16S ribosomal DNA fragments.Applied and environmental microbiology · 1997Article
- DNA rehybridization during PCR: the 'Cot effect' and its consequences.Nucleic acids research · 1996Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
Arbitrarily primed PCR fingerprinting of RNA and differential display resolved on an acrylamide gel has been extensively used to detect differentially expressed RNAs. However, after a differentially amplified product is detected the next steps are labor-intensive: a small portion of the fingerprinting gel that contains the differentially amplified product is cut out, reamplified and the correct product is determined, typically by cloning and sequencing what is often a mixture of products of similar size. Here we use a native acrylamide gel to separate DNAs in the reamplified mixture based on single-stranded conformation polymorphisms. Reamplifications are performed for the region carrying the differentially amplified product and a corresponding region from an adjacent lane where the product is less prominent or not visible. Denaturation of the reamplified DNA followed by side-by-side comparison on an SSCP gel allows the classification of reamplified material into (i) those that can be directly cloned because the differentially amplified product is relatively pure, (ii) those that need to be reamplified from the SSCP gel before cloning and (iii) those that are too complex for further study. This screen should save considerable effort now wasted on directly cloning unsuitable products from RNA fingerprinting experiments. An example is presented of cloning a gene differentially expressed in Trypanosoma brucei life cycle.
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