BIOB 375 1st Edition Lecture 31 Outline of Last Lecture II. ReviewOutline of Current Lecture III. DNA TechnologyIV. PCRCurrent LectureDNA TechnologyDNA technology includes:- DNA marker developmento To predict phenotype- Combining one DNA molecule with other DNA molecules- Gene amplificationDNA Marker development- To reveal polymorphisms between alleleso Polymorphisms: Difference at DNA sequence- Purpose of developing DNA markers: To make the polymorphisms visible, to predict phenotypes that require further testing. - Types of variations (between alleles):o Substitutiono Insertion/deletion - Two major strategies for detecting polymorphisms at DNA level:o PCR basedo Hybridization basedType of mutation: Base substitution mutationType of marker: Single nucleotide polymorphism (SNP)Type of mutation: Insertion/deletionType of marker: Simple Sequence repeat (SSR) & Sequence tagged site (STS)PCR (Polymerase Chain Reaction) based:- SNP: single nucleotide polymorphism- SSR: simple sequence repeat- STS: sequence tagged siteCombining one DNA molecule with other DNA molecules:Step 1: Cutting DNA by restriction enzymes, or mechanical forceStep 2: Joining DNA by ligaseA restriction enzyme (or restriction endonuclease) is an enzyme that cuts double-stranded DNA. The term restriction comes from the fact that these enzymes were discovered in E. coli strains that appeared to be restriction the infection by certain bacteriophages. There are two features about restriction enzymes:1. Recognizing special sequence2. Cutting sequence in a particular
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