Transcript REVIEW OF MOLECULAR GENETICS - Pascack Valley Regional

REVIEW OF MOLECULAR
GENETICS
DR. EDELBERG
Genes, DNA, & Chromosomes
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Anatomy of DNA
Gene anatomy-simple
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Anatomy of a Gene
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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
Genomic
Library
Construction
of cDNA
Library
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
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)
Synthesis of cDNA from mRNA
Cloning DNA into vector
Cloning into vectordetailed
Cloned into multiple
cloning site
Plasmid Characteristics
Ori
Selectable
Marker
3.6 Kb
Color screen
MCS
Circular DNA
Color
Screen
Transformation
Over night