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Molecular techniques in plant breeding, Cantho september 2006. 2. DAY 2: Methods for the analysis of DNA polymorphisms Chapter 4: Recombinant DNA, restriction enzymes, cloning, transformation 21 Chapter 5: Hybridisation, RFLP, micro-array

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    1. Molecular techniques in plant breeding, Cantho september 2006 1 Contents

    2. Molecular techniques in plant breeding, Cantho september 2006 2 Contents

    3. Molecular techniques in plant breeding, Cantho september 2006 3 Contents

    4. Molecular techniques in plant breeding, Cantho september 2006 4 Contents

    5. Molecular techniques in plant breeding, Cantho september 2006 5 Chapter 1: Genes and genomes DNA structure Genomes Protein structure Gene structure and expression

    6. Molecular techniques in plant breeding, Cantho september 2006 6 Cells and DNA

    7. Molecular techniques in plant breeding, Cantho september 2006 7 Chromosomes and DNA

    8. Molecular techniques in plant breeding, Cantho september 2006 8 DNA: Chemical Composition Two types of nitrogen-containing bases comprise the chemical structure of DNA: - purines = adenine and guanine, A & G - pyrimidines = thymine and cytosine, T & C

    9. Molecular techniques in plant breeding, Cantho september 2006 9 DNA: Chemical Composition Nucleotides = building blocks of DNA = phosphate + sugar + base Nucleoside = sugar +base Sugar = 5 carbon deoxyribose Phosphodiester bonds link sugar molecules to phosphate groups

    10. Molecular techniques in plant breeding, Cantho september 2006 10

    11. Molecular techniques in plant breeding, Cantho september 2006 11 DNA: Chemical Structure DNA = polynucleotide = a chain of bases Orientation of sugar-phosphate linkages = 5’ to 3’ as the phosphate attached to the 5’ carbon of one sugar is linked to the 3’ carbon of the next sugar Purine and pyrimidine bases are linked to the 1’ carbon of sugar

    12. Molecular techniques in plant breeding, Cantho september 2006 12

    13. Molecular techniques in plant breeding, Cantho september 2006 13 DNA: Chemical Structure DNA consists of two polynucleotide chains which run 5’ to 3’ in opposite directions = antiparallel DNA chains are held together by hydrogen bonds between bases DNA bases pair by Chargaff’s rules: - Adenine (A) pairs with Thymine (T) by 2 H-bounds - Guanine (G) pairs with Cytosine (C) by 3 bounds

    14. Molecular techniques in plant breeding, Cantho september 2006 14

    15. Molecular techniques in plant breeding, Cantho september 2006 15 DNA Sequence: convention 5’ to 3’end, one strand (because other strand is complementary and therefore known also)

    16. Molecular techniques in plant breeding, Cantho september 2006 16 Genome Size Complex organisms have large genomes = genetic contents of a cell Genomic size increases with evolutionary complexity (in general!) Size of DNA is measured in kb = kilobase pairs Size of large genomes is measured in Mb = megabase pairs

    17. Molecular techniques in plant breeding, Cantho september 2006 17 Genes and genomes Gene = unique sequence of DNA bases, coding for a protein Alleles = different variants of a gene in different organisms or on the homologous chromosomes

    18. Molecular techniques in plant breeding, Cantho september 2006 18 Genes and genomes

    19. Molecular techniques in plant breeding, Cantho september 2006 19

    20. 20 Chromosomes of a human being: 2 copies of each chromosome, ordered by size on a slide

    21. Molecular techniques in plant breeding, Cantho september 2006 21

    22. Molecular techniques in plant breeding, Cantho september 2006 22

    23. Molecular techniques in plant breeding, Cantho september 2006 23

    24. Molecular techniques in plant breeding, Cantho september 2006 24 Mitochondrial genomes Human: 16 kb Yeast: 84 kb Corn: 570 kb Genes for: rRNA, tRNA proteins for ATP synthesis

    25. Molecular techniques in plant breeding, Cantho september 2006 25 Chloroplast genomes 120 – 200 kb Genes voor: rRNA and tRNA RNA polymerase ca. 120 proteins for photosynthesis

    26. Molecular techniques in plant breeding, Cantho september 2006 26 Maternal inheritance of mitochondrial DNA

    27. Molecular techniques in plant breeding, Cantho september 2006 27 Organel DNA for tracing back

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