Application of inter simple sequence repeat (ISSR) markers to plant genetics

Godwin, ID, AitkenEAB and SmithLW (1997) Application of inter simple sequence repeat (ISSR) markers to plant genetics. Electrophoresis, 18 9: 1524-1528.


Author Godwin, ID
AitkenEAB
SmithLW
Title Application of inter simple sequence repeat (ISSR) markers to plant genetics
Journal name Electrophoresis   Check publisher's open access policy
ISSN 0173-0835
Publication date 1997
Sub-type Article (original research)
DOI 10.1002/elps.1150180906
Volume 18
Issue 9
Start page 1524
End page 1528
Total pages 5
Language eng
Abstract Microsatellites or simple sequence repeats (SSRs) are ubiquitous in eukaryotic genomes. Single-locus SSR markers have been developed for a number of species, although there is a major bottleneck in developing SSR markers whereby flanking sequences must be known to design 5'-anchors for polymerase chain reaction (PCR) primers. Inter SSR (ISSR) fingerprinting was developed such that no sequence knowledge was required. Primers based on a repeat sequence, such as (CA)(n), can be made with a degenerate 3'-anchor, such as (CA)(8)RG or (AGC)(6)TY. The resultant PCR reaction amplifies the sequence between two SSRs, yielding a multilocus marker system useful for fingerprinting, diversity analysis and genome mapping. PCR products are radiolabelled with P-32 or P-33 via end-labelling or PCR incorporation, and separated on a polyacrylamide sequencing gel prior to autoradiographic visualisation. A typical reaction yields 20-100 bands per lane depending on the species and primer. We have used ISSR fingerprinting in a number of plant species, and report here some results on two important tropical species, sorghum and banana. Previous investigators have demonstrated that ISSR analysis usually detects a higher level of polymorphism than that detected with restriction fragment length polymorphism (RFLP) or random amplified polymorphic DNA (RAPD) analyses. Our data indicate that this is not a result of greater polymorphism genetically, but rather technical reasons related to the detection methodology used for ISSR analysis.
Keyword Biochemical Research Methods
Chemistry, Analytical
Dna Markers
Genome Mapping
Genetic Diversity
Banana
Sorghum
Dna Markers
Polymorphisms
Amplification
Abundance
Diversity
Microsatellites
Identification
Location
Wheat
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Unknown

Document type: Journal Article
Sub-type: Article (original research)
Collection: School of Biological Sciences Publications
 
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Created: Mon, 13 Aug 2007, 16:58:50 EST