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Chapter 2

Chapter 2. DNA Biology Review. ©2002 Academic Press. chromosome. cell nucleus. Double stranded DNA molecule. Target Region for PCR. Individual nucleotides. DNA in the Cell. ©2002 Academic Press. Phosphate. Base. Sugar. Basic Components of Nucleic Acids. 5’end | Phosphate |

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Chapter 2

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  1. Chapter 2 DNA Biology Review ©2002 Academic Press

  2. chromosome cell nucleus Double stranded DNA molecule Target Region for PCR Individual nucleotides DNA in the Cell ©2002 Academic Press

  3. Phosphate Base Sugar Basic Components of Nucleic Acids 5’end | Phosphate | Sugar—Base… | Phosphate | Sugar—Base… | 3’end ©2002 Academic Press

  4. Hydrogen bonds 5’ 3’ 5’ A = T 3’ A = T G  C hybridized strands G  C A = T T = A Phosphate-sugar backbone T = A C  G C  G A = T A T T = A A T G C C  G C  G G C denatured strands C G 5’ 3’ G  C 3’ 5’ Base Pairing of DNA Strands ©2002 Academic Press

  5. Chromosome 12 telomere p (short arm) 12p3 Band 3 centromere q (long arm) 12q5 Band 5 telomere Basic Chromosome Structure ©2002 Academic Press

  6. http://www.ncbi.nlm.nih.gov/genome/guide/ 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 X Y Sex-chromosomes Human Genome Contains 23 Pairs of Chromosomes ©2002 Academic Press

  7. TYPES OF DNA MARKERS (GATA)(GATA)(GATA)(GATA) • Length Variation • short tandem repeats (STRs) • microsatellites • simple sequence repeats (SSRs) • minisatellites (VNTRs) • Sequence Variation • single nucleotide polymorphisms (SNPs) • insertions/deletions (CA)(CA)(CA)(CA(CA)(CA) (G/A) ©2002 Academic Press

  8. Family Inheritance of STR Alleles (D13S317) PCR product size (bp) 11 14 Father 12 14 Child #1 8 14 Child #2 11 12 Child #3 8 12 Mother ©2002 AP, Elsevier

  9. Time required to obtain results 6-8 weeks with radioactive probes; ~1 week with chemiluminescent probes 1-2 days Characteristic RFLP Methods PCR Methods Amount of DNA needed 50-500 ng 0.1-1 ng Condition of DNA needed high molecular weight, intact DNA may be highly degraded Capable of handling sample mixtures Yes (single locus probes) Yes Allele identification Binning required Discrete alleles obtained Power of Discrimination ~1 in 1 billion with 6 loci ~1 in 1 billion with 8-13 loci (requires more loci) ©2002 AP, Elsevier Comparison of RFLP and PCR

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