The primary objectives of these laboratory exercises are to familiarize advanced undergraduate and graduate students (and instructors) with the general concepts, techniques, and uses of DNA fingerprinting and to remove some of the perceived mystique underlying molecular genetics. The technique of DNA amplification fingerprinting (DAF) is partitioned into four independent laboratory exercises that include DNA isolation, DNA amplification, gel electrophoresis and silver staining, and data collection and analysis. Although the DNA amplification and gel electrophoresis exercises are emphasized, very detailed and easy-to-follow instructions and protocols are provided for all aspects of the DNA fingerprinting process. These exercises, or similar ones, have been successfully completed on the first attempt by several classes of novice graduate students and other researchers.
R.N. Trigiano and G. Caetano-Anollés
David Jesús Gil-Ariza, Iraida Amaya, José Manuel López-Aranda, José Federico Sánchez-Sevilla, Miguel Ángel Botella and Victoriano Valpuesta
accessions cover both a wide range of temporal and geographical origins (since the 18th century until today) and a representative group of pedigree-related recent cultivars, which were selected to test the power of SSR markers in cultivar identification
Imen Rekik, Amelia Salimonti, Naziha Grati Kamoun, Innocenzo Muzzalupo, Oliver Lepais, Sophie Gerber, Enzo Perri and Ahmed Rebai
polymerase chain reaction (PCR) and are highly reproducible among different laboratories. Almost all reports of cultivar identification using SSRs assessed differences in lengths of amplified alleles ( Bandelj et al., 2002 , 2004 ; Belaj et al., 2004
Àngel Fernández i Martí, José M. Alonso, María T. Espiau, María J. Rubio-Cabetas and Rafel Socias i Company
( Prunus persica L.) ( Messeguer et al., 1987 ) and almond ( Cerezo et al., 1989 ; Hauagge et al., 1987 ). Later, DNA markers were introduced for cultivar identification, such as restriction fragment length polymorphisms (RFLPs) in apricot [ Prunus
Shuang Jiang, Haishan An, Xiaoqing Wang, Chunhui Shi, Jun Luo and Yuanwen Teng
cultivar identification ( Kimura et al., 2002 ), genetic diversity studies ( Bao et al., 2007 ), and quantitative trait locus mapping ( Perchepied et al., 2015 ; Yamamoto et al., 2014 ). Compared with other molecular markers, SSR markers provide a number
Shengrui Yao, Junxin Huang and Robert Heyduck
for cultivar identification. Table 2. Jujube flower diameter of different cultivars at New Mexico State University’s Sustainable Agriculture Science Center at Alcalde, New Mexico. Flower blooming type. Jujube cultivars can be classified as two blooming
Sriyani Rajapakse, Mark Hubbard, Albert Abbott, Robert Ballard and John Kelly
Restriction Fragment Length Polymorphisms (RFLPs) were investigated in rose cultivars as a means of reliable cultivar identification. A random genomic DNA library was generated by shotgun cloning HindIII digested fragments of DNA extracted from rose cultivar Confection into pUC8 plasmid of Escherichia coli strain JM 83. Compared to genomic clones carrying low or highly repeated sequences, clones with moderately repeated sequences were most effective in cultivar identification. These clones were identified by hybridizing rose DNA fragments from the library with genomic DNA from `Confection'. Clones with moderately repeated copy sequences were used as probes to detect the presence of RFLPs by Southern hybridization of EcoRI digested genomic DNA of various rose cultivars. Several of these probes have revealed RFLPs useful in cultivar identification. By using a combination of two or more of these probes most of the rose cultivars compared at this time can be identified. A dichotomous key useful in identification of rose cultivars was prepared from RFLPs displayed by 3A9 probe.
Gayle M. Volk, Adam D. Henk and Christopher M. Richards
We thank Walter Lyons, Barbara Hellier, and Tom Cloud for providing garlic samples and assisting with cultivar identification. We appreciate the technical assistance provided by Jackie Harris and Pat Reeves. Mention of trade names or commercial
David B. Rubino
Segregating progenies from controlled pollinations of Eustoma grandiflorum Griseb. were investigated to determine the inheritance of diaphorase (DIA) and glucoses-phosphate isomerase (GPI) isozymes. Phenotypic data supported the hypotheses that DIA1 is tetrameric and is controlled by a single locus with two alleles (Dia1-1 and Dia1-2) and that GPI1 is dimeric and also is controlled by a single locus with two alleles (Gpi1-1 and Gpi1-2). Examination of isozyme phenotypes for over 70 cultivars of E. grandiflorum revealed polymorphism for DIA1 and GPI1. These isozymes may be useful for marker-assisted selection and cultivar identification.
J.I. Hormaza, L. Dollo and V.S. Polito
The Random Amplified Polymorphic DNA (RAPD) technique was used to characterize 15 cultivars of pistachio (Pistacia vera L.). A total of 37 polymorphic markers were considered in this study. Each cultivar exhibited a unique molecular phenotype and, as a consequence, can be uniquely fingerprinted. A similarity and cluster analysis based on the amplified fragments produced two distinct groups which are consistent with the known geographical origin of the cultivars. Our results suggest that RAPD analysis can provide a new alternative for cultivar identification and classification of pistachio.