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Specific Primer Pairs (specific + primer_pair)

Distribution by Scientific Domains
Chemistry50%

Selected Abstracts

Identification of S -alleles using polymerase chain reaction-cleaved amplified polymorphic sequence of the S -locus receptor kinase in inbreeding lines of Brassica oleracea

PLANT BREEDING, Issue 3 2002
J. I. Park
Abstract Identification and DNA polymorphism of the S -locus receptor kinasegene (SRK) was analysed by pollen tube tests, polymerase chain reaction-cleaved amplified polymorphic sequence (PCR-CAPS) and nucleotide sequencing. SRK -specific primers that can distinguish class and class II S haplotypes amplified single DNA fragments of 900-1050 bp. The DNA fragments of 22 inbred lines amplified with a class SRK -specific primer pair determined seven types with HinfI and EcoRII. In addition, the DNA fragments of 17 inbred lines amplified with a class II SRK -specific primer pair determined three types with Hinf1. Nucleotide sequencing of the DNA fragments amplified from 10S haplotypes showed that exons of the 3,-end in SRK are highly conserved, and that there is much variation of the introns, which produced polymorphism of the band pattern in PCR-CAPS profiles. The S haplotypes of the plants were determined by restriction analysis of PCR products and agreed with results based on pollen tube growth tests. The PCR-CAPS analysis using specific primer pairs of SRK is considered to be useful for S allele identification in breeding programmes. [source]

Use of specific PCR-based molecular markers for discrimination, rapid analysis of purity and identification of six fragrant rice varieties

INTERNATIONAL JOURNAL OF FOOD SCIENCE & TECHNOLOGY, Issue 10 2009
Ho Ky Quang Minh
Summary Fragrant rice is high in quality and hence price. Therefore, fragrant rice is often mixed with non-fragrant or low grade rice. The quality of fragrant rice depends on many factors such as variety, region in which it is cultivated, handling practices, etc. In this study, two specific primer pairs, including external sense primer, external antisense primer, internal non-fragrant sense primer and internal fragrant antisense primer were used to differentiate fragrant and non-fragrant rice based on the detection of 8-bp deletion in betaine aldehyde dehydrogenase 2 gene. The purity of the fragrant rice can be detected at 1% and 2% level of adulteration with homogeneous and heterogeneous non-fragrant rice varieties, respectively, using only one primer pair (internal non-fragrant sense primer and external antisense primer). Furthermore, six particular fragrant rice varieties were identified using specific RM 1 microsatellite markers. Further analysis should be conducted to verify the discriminating power of this method of analysis for the traceability of fragrant rice varieties. [source]

SIMULTANEOUS DETECTION OF LISTERIA MONOCYTOGENES, STAPHYLOCOCCUS AUREUS, SALMONELLA ENTERICA AND ESCHERICHIA COLI O157:H7 IN FOOD SAMPLES USING MULTIPLEX PCR METHOD

JOURNAL OF FOOD SAFETY, Issue 3 2009
D. ZHANG
ABSTRACT In this study, one multiplex polymerase chain reaction (MPCR) assay was developed for simultaneous detection of four foodborne pathogens, i.e., Listeria monocytogenes, Staphylococcus aureus, Salmonella enterica and Escherichia coli O157:H7. Five specific primer pairs were designed based on the nucleotide sequences of hemolysin gene (hly) of Listeria monocytogenes, thermostable nuclease gene (nuc) of Staphylococcus aureus, invasion gene (invA) of Salmonella enterica, shiga-like toxin gene (stx) and intimin gene (eae) of Escherichia coli O157:H7 in this assay. The specificity and sensitivity of the MPCR method were validated, and the limit of detection (LOD) of this method was about 10 copies. One cfu/mL each of these foodborne pathogens spiked in practical food samples, i.e., ground meat, beef, pork, fish, shrimp, cheese, canola leaf and cabbage, could be detected simultaneously after 24 h enrichment at a rate of 87.5%, indicating that the established MPCR detection method was effective and suitable for practical use. PRACTICAL APPLICATIONS This study presents a quick and effective identification method to simultaneous monitor four foodborne pathogens in food samples. The specificity and sensitivity of this method can be used to unambiguously identify these four foodborne pathogens in practical food samples based on the species-specific genes. Therefore, this detection method is applicable for surveillance measures of these four foodborne pathogens in the food production chain. [source]

Identification of S -alleles using polymerase chain reaction-cleaved amplified polymorphic sequence of the S -locus receptor kinase in inbreeding lines of Brassica oleracea

PLANT BREEDING, Issue 3 2002
J. I. Park
Abstract Identification and DNA polymorphism of the S -locus receptor kinasegene (SRK) was analysed by pollen tube tests, polymerase chain reaction-cleaved amplified polymorphic sequence (PCR-CAPS) and nucleotide sequencing. SRK -specific primers that can distinguish class and class II S haplotypes amplified single DNA fragments of 900-1050 bp. The DNA fragments of 22 inbred lines amplified with a class SRK -specific primer pair determined seven types with HinfI and EcoRII. In addition, the DNA fragments of 17 inbred lines amplified with a class II SRK -specific primer pair determined three types with Hinf1. Nucleotide sequencing of the DNA fragments amplified from 10S haplotypes showed that exons of the 3,-end in SRK are highly conserved, and that there is much variation of the introns, which produced polymorphism of the band pattern in PCR-CAPS profiles. The S haplotypes of the plants were determined by restriction analysis of PCR products and agreed with results based on pollen tube growth tests. The PCR-CAPS analysis using specific primer pairs of SRK is considered to be useful for S allele identification in breeding programmes. [source]