Journal article
Target region amplification polymorphism: A novel marker technique for plant genotyping
Plant molecular biology reporter, Vol.21(3), pp.289-294
09/2003
Handle:
https://hdl.handle.net/2376/112072
Abstract
The advent of large-scale DNA sequencing technology has generated a tremendous amount of sequence information for many important organisms. We have developed a rapid and efficient PCR-based technique, which uses bioinformatics tools and expressed sequence tag (EST) database information to generate polymorphic markers around targeted candidate gene sequences. This target region amplification polymorphism (TRAP) technique uses 2 primers of 18 nucleotides to generate markers. One of the primers, the fixed primer, is designed from the targeted EST sequence in the database; the second primer, the arbitrary primer, is an arbitrary sequence with either an AT-or GC-rich core to anneal with an intron or exon, respectively. PCR amplification is run for the first 5 cycles with an annealing temperature of 35°C, followed by 35 cycles with an annealing temperature of 50°C. For different plant species, each PCR reaction can generate as many as 50 scorable fragments with sizes ranging from 50–900 bp when separated on a 6.5% polyacrylamide sequencing gel. The TRAP technique should be useful in genotyping germplasm collections and in tagging genes governing desirable agronomic traits of crop plants.
Metrics
24 Record Views
Details
- Title
- Target region amplification polymorphism: A novel marker technique for plant genotyping
- Creators
- Jinguo Hu - Northern Crop Science Laboratory USDA-ARS 58105 Fargo NDBrady Vick - Northern Crop Science Laboratory USDA-ARS 58105 Fargo ND
- Publication Details
- Plant molecular biology reporter, Vol.21(3), pp.289-294
- Academic Unit
- Agricultural, Human, and Natural Resource Sciences, College of
- Publisher
- Springer-Verlag; New York
- Identifiers
- 99900547790701842
- Language
- English
- Resource Type
- Journal article