Optimizing the generation of random amplified polymorphic DNAs in chrysanthemum

Wolff, Kirsten; Schoen, Eric D.; Rijn, J. Peters-van

doi:10.1007/bf00211058

Cited by 58 publications

(34 citation statements)

References 7 publications

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“…In a previous experiment the reaction conditions were optimized (Wolff et at., 1993). Reactions were performed in a volume of 50 1u1 containing 20 mM Tris-HCI, pH 8.3, 50 mivi KC1, 3mM MgCl2, 0.001 per cent gelatin, 100 UM each of dATP, dCTP, cGTP and dTTP, 0.2 1UM primer, 25 ng of chrysanthemum genomic DNA, and 1 U of Taq polymerase (Amplitaq, Perkin Elmer Cetus) using a Perkin Elmer 9600 thermal cycler.…”

Section: Materials and Methods P/ant Mater/at And Experimental Setupmentioning

confidence: 99%

Rapid detection of genetic variability in chrysanthemum (Dendranthema grandiflora Tzvelev) using random primers

1993

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Genetic variation in chrysanthemum (Dendranthema grandifiora) was studied using a recently developed technique generating Random Amplified Polymorphic DNAs (RAPDs). It appeared that variation between cultivars was high and that the cultivars used could be distinguished from each other by using only two different primers. A family of cultivars, derived from one original cultivar by vegetative propagation, had identical fragment patterns. Because of the high level of polymorphism and clonal stability RAPD fragments are useful for cultivar identification. Genetic variability among related Dendranthema species was too high to study genetic distances either among cultivars within chrysanthemum or among species related to chrysanthemum.

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Section: Materials and Methods P/ant Mater/at And Experimental Setupmentioning

confidence: 99%

Rapid detection of genetic variability in chrysanthemum (Dendranthema grandiflora Tzvelev) using random primers

1993

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“…Most of these factors have now been standardised within certain narrow limits and optimisation is normally not required, but caution is still required to avoid potential problems (Wolff et al 1993). However one factor, the concentration of MgCl 2 must be optimised for each combination of primer-template tested to maximise the generation of DNA products (Park and Kohel 1994).…”

Section: Rapd-pcr Profilesmentioning

confidence: 99%

Differences in genomic DNA extracted from bark and from wood of different zones in Robinia trees using RAPD-PCR

Filippis

Magel

1998

Trees

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Determination of genetic differences and levels of gene expression in mature and old tissues (e.g. wood) is often difficult based on morphological and anatomical characteristics, levels of metabolites or enzymatic activity. The use of molecular markers allows assessment of polymorphic (genetic) variation amongst individuals and between closely related species directly at the DNA level, but such techniques have not been generally applied to the bark and wood of mature trees. In this study we have applied the technique of random amplification of polymorphic DNA (RAPD) by the polymerase chain reaction (PCR) to analyse the relationship between the bark and variously aged wood zones of Robinia pseudoacacia. The use of micro polyacrylamide gel electrophoresis coupled to silver staining for DNA provided a quick, reliable and sensitive method of detecting polymorphisms. It was necessary to test a small number of ten-base synthetic oligonucleotide primers before arriving at a set of five which clearly identified post-transcriptional differences between bark, sapwood, transition zone and heartwood even in the one individual tree. The variability of the technique, and in particular the origin and quality of the DNA extracted was analysed. We demonstrated that the procedures and protocols developed are applicable to all tissue types tested from bark to the inner heartwood zones. Our results show that RAPD-PCR technology is a versatile and sensitive method of detecting genomic changes in trees.

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“…As the PCR amplification process is dependent upon many components and their interactions (Devos and Gale, 1992;Caetano-Anolles and Bassam, 1993;Wolf et al, 1993), it is important to specify a set of reaction conditions in order to obtain reproducible results for a given species. Sources of reliability lie in the purity of the template DNA, magnesium (Mg 2+ ) concentration, the choice of thermal-stable DNA polymerase and thermal cycler used in PCR amplification.…”

Section: Random Amplified Polymorphic Dna (Rapd)mentioning

confidence: 99%

Cassava Genetic Improvement

Ceballos¹,

Fregene²,

Carlos³

et al. 2007

Breeding Major Food Staples

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PREFACEThis Handbook is the result of a combined effort by several current and previous cassava researchers at CIAT to review and summarize the most important results of cassava research during the past 40 years. Most of the chapters are based on the various presentations during the Regional Cassava Training Course, held in Thailand from October 6 to 17, 2008. This course was organized upon the realization that many people that were actively involved with cassava research in the 1970s and 80s, both at CIAT and in national programs, have now retired or will soon do so, and that a whole new generation of cassava researchers are currently being hired to take their place.As cassava is now becoming a very important, and mostly industrial, crop in Asia, there are many new opportunities, but also a host of new problems and challenges. These include the appearance in Asia of new cassava diseases and pests; the decreasing availability and increasing cost of rural labor, resulting in the need for partial or complete mechanization of cassava production; the rapidly increasing demand for cassava roots for production of food, feed and fuel, and the unavailability in many countries of new land for any expansion of cassava area, thus requiring a rapid increase in cassava yields to increase supplies. This requires a renewed focus on cassava research for the development of new higher-yielding varieties and more sustainable production practices.While many cassava researchers in national programs in Asia received individual or group training at CIAT-Colombia during the 1970s and 80s, this training was greatly reduced during the following two decades due to funding limitations. Thus, the objective of the Regional Cassava Training Course in 2008 was to provide a new training opportunity for young scientists in Asian countries, and to hand over the knowledge and experience of the older cassava researchers, mostly from CIAT, to a new generation that will have to face the new challenges. Thus, the course was taught mostly by current or already retired CIAT cassava researchers, while the 60-plus participants of the course included cassava researchers from Cambodia, China, East Timor, India, Indonesia, Lao PDR, Malaysia, Myanmar, the Philippines, Thailand and Vietnam. The training course not only provided knowledge -from physiology and biotechnology to animal feeding and production of fuelethanol -but also an opportunity for people from different institutions and countries to get to know each other, which will greatly facilitate future collaboration.Course participants returned home with a CD containing the PowerPoint presentations of the course. However, it was felt that a more comprehensive review of all the topics covered was warranted, as this would provide more in-depth knowledge for those working in the various specialized fields. Most of this information is available in many refereed journals, in workshop proceedings and old CIAT annual reports, but many of these are now out of print or otherwise difficult to obtain. Thus, the Cas...

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Optimizing the generation of random amplified polymorphic DNAs in chrysanthemum

Cited by 58 publications

References 7 publications

Rapid detection of genetic variability in chrysanthemum (Dendranthema grandiflora Tzvelev) using random primers

Rapid detection of genetic variability in chrysanthemum (Dendranthema grandiflora Tzvelev) using random primers

Differences in genomic DNA extracted from bark and from wood of different zones in Robinia trees using RAPD-PCR

Cassava Genetic Improvement

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