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REVIEW OF MOLECULAR GENETICS

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REVIEW OF MOLECULAR GENETICS DR. EDELBERG Genes, DNA, & Chromosomes Anatomy of DNA Gene anatomy-simple Anatomy of a Gene DNA LIBRARIES Genomic Library Construction of ... – PowerPoint PPT presentation

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Title: REVIEW OF MOLECULAR GENETICS


1
REVIEW OF MOLECULAR GENETICS
  • DR. EDELBERG

2
Genes, DNA, Chromosomes
3
Anatomy of DNA
4
Gene anatomy-simple
5
Anatomy of a Gene
6
DNA LIBRARIES
  • DNA library - a random collection of DNA
    fragments from an organism cloned into a vector
  • Ideally contains at least one copy of every DNA
    sequence.
  • Easily maintained in the laboratory
  • Can be manipulated in various ways to facilitate
    the isolation of a DNA fragment of interest to a
    scientist.
  • Numerous types of libraries exist for various
    organisms - Genomic and cDNA

7
Genomic Library
8
Construction of cDNA Library
9
Genomic Vs. cDNA Library
  • Genomic Library
  • Promoters
  • Introns
  • Intergenic
  • Non-expressed genes
  • cDNA Library
  • Expressed genes
  • Transcription start sites
  • Open reading frames (ORFs)
  • Splice points

10
Extraction of mRNA
Collect and grind up animals in mild denaturing
solution
Spin out debris (Tissue, membranes, etc)
Treat with DNAse (removes DNA)
Treat with Phenol (removes protein)
11
Synthesis of cDNA from mRNA
12
Cloning DNA into vector
13
Cloning into vector-detailed
14
Cloned into multiple cloning site
15
Plasmid Characteristics
  • Ori
  • Selectable Marker
  • 3.6 Kb
  • Color screen
  • MCS
  • Circular DNA

16
Color Screen
17
Transformation
18
Over night
19
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