| Title: |
High-Resolution RT-PCR Analysis of Alternative Barley Transcripts |
| Authors: |
Simpson, Craig G.; Fuller, John; Rapazote-Flores, Paulo; Mayer, Claus Dieter; Calixto, Cristiane P. G.; Milne, Linda; Hedley, Pete E.; Booth, Clare; Waugh, Robbie; Brown, John W. S. |
| Contributors: |
Harwood, Wendy A. |
| Source: |
Simpson, C G, Fuller, J, Rapazote-Flores, P, Mayer, C D, Calixto, C P G, Milne, L, Hedley, P E, Booth, C, Waugh, R & Brown, J W S 2019, High-Resolution RT-PCR Analysis of Alternative Barley Transcripts. in W A Harwood (ed.), Barley : Methods and Protocols. vol. 1900, Methods in Molecular Biology, Humana Press, New York, pp. 269-281. https://doi.org/10.1007/978-1-4939-8944-7_17 |
| Publisher Information: |
Humana Press |
| Publication Year: |
2019 |
| Collection: |
Discovery - University of Dundee Online Publications |
| Subject Terms: |
Alternative splicing; HR RT-PCR; RNA-seq; /dk/atira/pure/subjectarea/asjc/1300/1312; name=Molecular Biology; /dk/atira/pure/subjectarea/asjc/1300/1311; name=Genetics |
| Description: |
Assembly of the barley genome and extensive use of RNA-seq has resulted in an abundance of gene expression data and the recognition of wide-scale production of alternatively spliced transcripts. Here, we describe in detail a high-resolution reverse transcription-PCR based panel (HR RT-PCR) that confirms the accuracy of alternatively spliced transcripts from RNA-seq and allows quantification of changes in the proportion of splice isoforms between different experimental conditions, time points, tissues, genotypes, ecotypes, and treatments. By validating a selection of barley genes, use of the panel gives confidence or otherwise to the genome-wide global changes in alternatively spliced transcripts reported by RNA-seq. This simple assay can readily be applied to perform detailed transcript isoform analysis for any gene in any species. |
| Document Type: |
book part |
| File Description: |
application/pdf |
| Language: |
English |
| ISBN: |
978-1-4939-8942-3; 1-4939-8942-1 |
| Relation: |
info:eu-repo/semantics/altIdentifier/pmid/30460571; info:eu-repo/semantics/altIdentifier/isbn/9781493989423; urn:ISBN:9781493989423 |
| DOI: |
10.1007/978-1-4939-8944-7_17 |
| Availability: |
https://discovery.dundee.ac.uk/en/publications/0e106293-4378-4d03-8300-1381e71e94e5; https://doi.org/10.1007/978-1-4939-8944-7_17; https://discovery.dundee.ac.uk/ws/files/29490735/2016_Methods_Mol_Biol_AS_CS_Nov_2016_edits_CS_1_.pdf; https://www.scopus.com/pages/publications/85056802477 |
| Rights: |
info:eu-repo/semantics/openAccess |
| Accession Number: |
edsbas.3B4433F1 |
| Database: |
BASE |