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Lookup NU author(s): Dr Mohammed Ali, Professor John IsaacsORCiD
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Differential display is one of the simplest techniques for discovering novel transcripts when comparing gene expression in biological systems. The method can be carried out on small amounts of total RNA and permits the simultaneous comparison of multiple independent samples in a single experiment. The methodology is versatile in that it allows the researcher to adapt the existing protocol by varying the selection of oligonucleotide primers used in the PCR. The putative differentials, which are isolated from the polyacrylamide gels as cDNA fragments of approx 100 to 500 bases in size, can be instantly recognized after sequencing by searching the nucleotide databases. However, one of the drawbacks is the isolation of false positives and hence the need to confirm the results of the screen by another method. Once the true differentials have been identified, further downstream work is also required to recognize which splice variant of the transcript gives rise to the expression differences and whether the gene expression results are corroborated at the protein level.
Author(s): Ali M, Isaacs JD
Editor(s): Cope, AP
Publication type: Book Chapter
Publication status: Published
Book Title: Arthritis Research
Year: 2007
Volume: 136
Pages: 329-47
Series Title: Methods in Molecular Medicine
Publisher: Humana Press
Place Published: US
URL: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17983158
DOI: 10.1007/978-1-59745-402-5_23
Notes: ISSN: 1543-1894 (Print) 1940-6037 (Online) Online ISBN: 9781597454025
Library holdings: Search Newcastle University Library for this item
ISBN: 9781588299185