Zea mays L. - VAAD

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CPVO-TP/002/3 FinalEnglishDate: 11/03/2010Characteristics Examples Note48. Allele expression atlocus Pgi 1 Genotype 4/4 A239 1QL Genotype 5/5 A632 2Genotype 4/5 Artist 349. Allele expression atlocus Acp1 Genotype 2/2 F 2 1PQ Genotype 2/3 Azur 2Genotype 3/3 A 239 3Genotype 4/6 Contessa 4Genotype 4/4 A 632 5Genotype 6/6 F 1444 6Genotype 2/4 Occitan 7Genotype 2/6 8Genotype 3/4 Marshall 9Genotype 3/6 1050. Allele expression atlocus Dia 1 Genotype 8/8 F 2 1QL Genotype 12/12 CO 158 2Genotype 8/12 Bastion 351. Allele expression atlocus Dia2 Genotype 4/4 F 2 1QL Genotype 6/6 34 M 838 2Genotype 4/6 31 N 6 352. Allele expression atlocus Adh 1 Genotype 4/4 F 1444 1QL Genotype 6/6 F 2 2Genotype 4/6 Bristol 330
CPVO-TP/002/3 FinalEnglishDate: 11/03/2010Part IIIDescription of the SGE Method for theAnalysis of Isozymes from <strong>Zea</strong> <strong>mays</strong> L.1. Number of coleoptiles per test- for checking formula: at least 20 coleoptiles of each inbred line2 coleoptiles of single-cross hybrids6 coleoptiles of three-way cross hybrids- for distinctness, uniformity and stability test: at least 20 coleoptiles for inbredlines, hybrids and open-pollinated varieties.2. Apparatus and equipmentAny suitable horizontal electrophoresis system can be used, provided that the gels can be kept at 4°C. A gelthickness of 10 mm is recommended. The power supply used should be capable of delivering constant voltageoutput.3. ChemicalsAll chemicals should be of 'Analytical Reagent' grade or better.3.1 Chemicals for enzyme extractionL-Ascorbic acidL-Ascorbic acid Na saltSucrose3.2 Chemicals for electrophoresisBromophenol blueCitric acid monohydrateL-HistidineStarch hydrolyzed, for electrophoresis,)3.3 Chemicals for staining enzymesAcetic acid glacial2,6-Dichlorophenol-indophenol Na saltEthanolEthylenediamine tetra-acetic acid Na2 Salt (EDTA)Fast Garnet GBC saltD-Fructose 6-phosphate Na2 saltGlucose 1-phosphate dehydrogenase (Serva 22820 or 22822 or Sigma G5885)Hydrochloric acid (HCl)DL-Isocitric acid Na3 saltMagnesium chloride hexahydrateDL-Malic acidDimethylthiazol diphenyl tetrazolium (MTT)β -Nicotinamide adenine dinucleotide (NAD)β -Nicotinamide adenine dinucleotide reduced (NADH)β -Nicotinamide adenine dinucleotide phosphate (NADP)Nitro-blue tetrazolium (NBT)Sodium hydroxide (NaOH)1-Naphtyl acid phosphate6-phosphogluconic acid Na3 salt dihydratePhenazine methosulfate (PMS)Polyvinylpyrrolidone 40 (PVP-40)Sodium acetate trihydrateTris-(hydroxymethyl) aminomethane (Tris)31
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