REVIEW OF MOLECULAR GENETICS DR. EDELBERG

Genes, DNA, & Chromosomes

Anatomy of DNA

Gene anatomy-simple

Anatomy of a Gene

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  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 Genomic Library Promoters Introns Intergenic Non-expressed genes cDNA Library Expressed genes Transcription start sites Open reading frames (ORFs) Splice points 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 vector- detailed

Cloned into multiple cloning site

Plasmid Characteristics FOri FSelectable Marker F3.6 Kb FColor screen FMCS FCircular DNA FOri FSelectable Marker F3.6 Kb FColor screen FMCS FCircular DNA

Color Screen

Transformation

Over night