MersV1, Istex, Corpus, bibRecord, 000195

Large-scale, cost-effective screening of PCR products in marker-assisted selection applications

Identifieur interne : 000195 ( Istex/Corpus ); précédent : 000194; suivant : 000196

Large-scale, cost-effective screening of PCR products in marker-assisted selection applications

Auteurs : W. K. Gu ; N. F. Weeden ; J. Yu ; D. H. Wallace

Source :

Theoretical and Applied Genetics [ 0040-5752 ] ; 1995-08-01.

RBID : ISTEX:4FB1CAFEC7E123A3BABD4C9BB72E52DDA4BB793D

English descriptors

KwdEn :
- Allele-specific associated primers, Common bean, Electrophoresis, Pea, RAPDs.
Teeft :
- Allele, Allele specificity, Amplification, Annealing, Annealing temperature, Annealing temperatures, Appl, Appropriate allele, Arbitrary primers, Asap, Asap primers, Asaps, Bromide, Clone, Common bean, Cornell university, Correct size, Cycle parameters, Detection methods, Detection steps, Different plant, Direct measurement, Direct staining, Double helix, Enation mosaic virus, Error rate, Ethidium, Ethidium bromide, Extraction procedure, Fragment, Free nucleotides, Genet, Genetic markers, Genotype, Identical size, Large numbers, Leaf tissue, Marker, Marker detection, Marker sequence, Michelmore, Nucleic acids, Phaseolus vulgaris, Photoperiod, Photoperiod genes, Pisum sativum, Primer, Rapd, Rapd fragment, Rapd markers, Reaction mixture, Reaction product, Second amplification, Selection applications, Shorter primers, Simple procedures, Specific primers, Theor, Theor appl genet, Thermal cycler, Weak fluorescence, Weeden.

Abstract

Abstract: A simple, PCR-based method has been developed for the rapid genotyping of large numbers of samples. The method involves a alkaline extraction of DNA from plant tissue using a slight modification of the procedure of Wang et al. (Nucleic Acids Res 21:4153–4154, 1993). Template DNA is amplified using allelespecific associated primers (ASAPs) which, at stringent annealing temperatures, generate only a single DNA fragment and only in those individuals possessing the appropriate allele. This approach eliminates the need to separate amplified DNA fragments by electrophoresis. Instead, samples processing the appropriate allele are identified by direct staining of DNA with ethidium bromide. Total technician time required for extraction, amplification and detection of 96 samples is about 4 h, and this time requirement can be reduced by automation. Excluding labor, cost per sample is less than $0.40. The method is tested using the codominant isozyme marker, alcohol dehydrogenase (Adh-1) gene in pea (Pisum sativum), and applied to the screening of photoperiod genes in common bean (Phaseolus vulgaris L.).

Url:

https://api.istex.fr/ark:/67375/1BB-J9N95TTW-D/fulltext.pdf

DOI: 10.1007/BF00222974

Links to Exploration step

ISTEX:4FB1CAFEC7E123A3BABD4C9BB72E52DDA4BB793D

Le document en format XML

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<Para>A simple, PCR-based method has been developed for the rapid genotyping of large numbers of samples. The method involves a alkaline extraction of DNA from plant tissue using a slight modification of the procedure of Wang et al. (Nucleic Acids Res 21:4153–4154, 1993). Template DNA is amplified using allelespecific associated primers (ASAPs) which, at stringent annealing temperatures, generate only a single DNA fragment and only in those individuals possessing the appropriate allele. This approach eliminates the need to separate amplified DNA fragments by electrophoresis. Instead, samples processing the appropriate allele are identified by direct staining of DNA with ethidium bromide. Total technician time required for extraction, amplification and detection of 96 samples is about 4 h, and this time requirement can be reduced by automation. Excluding labor, cost per sample is less than $0.40. The method is tested using the codominant isozyme marker, alcohol dehydrogenase (<Emphasis Type="Italic">Adh-1</Emphasis>
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<abstract lang="en">Abstract: A simple, PCR-based method has been developed for the rapid genotyping of large numbers of samples. The method involves a alkaline extraction of DNA from plant tissue using a slight modification of the procedure of Wang et al. (Nucleic Acids Res 21:4153–4154, 1993). Template DNA is amplified using allelespecific associated primers (ASAPs) which, at stringent annealing temperatures, generate only a single DNA fragment and only in those individuals possessing the appropriate allele. This approach eliminates the need to separate amplified DNA fragments by electrophoresis. Instead, samples processing the appropriate allele are identified by direct staining of DNA with ethidium bromide. Total technician time required for extraction, amplification and detection of 96 samples is about 4 h, and this time requirement can be reduced by automation. Excluding labor, cost per sample is less than $0.40. The method is tested using the codominant isozyme marker, alcohol dehydrogenase (Adh-1) gene in pea (Pisum sativum), and applied to the screening of photoperiod genes in common bean (Phaseolus vulgaris L.).</abstract>
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Large-scale, cost-effective screening of PCR products in marker-assisted selection applications

Large-scale, cost-effective screening of PCR products in marker-assisted selection applications

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