This preview shows pages 1–3. Sign up to view the full content.
This preview has intentionally blurred sections. Sign up to view the full version.View Full Document
Unformatted text preview: Chapter 20 DNA Technology VOCABULARY Polymerase Chain Reaction (PCR), DNA replication, primers, dNTPs Thermal cycling: denaturation, annealing, elongation Taq DNA polymerase DNA sequencing, ddNTPs, dyes, acrylamide FIGURES: 20.7; 20.12; 20.14 CONCEPTS PCR uses an enzymatic reaction and the ability of nucleotides to form hydrogen bonds (A:T, C:G) to amplify a gene (Sanger) DNA sequencing uses an enzymatic reaction and the ability of nucleotides to form hydrogen bonds to determine a sequence of nucleotides NOTE: YOU ARE NO LONGER RESPONSIBLE FOR MICROARRAYS All take advantage of biochemical properties to manipulate DNA PCR amplifies a specific gene using an enzymatic reaction and the tendency of A to bind T and C to bind G PCR is very similar to DNA replication , but in a test tube! You need: DNA polymerase : enzyme to perform the reaction: single enzyme reaction! dNTPs : building blocks of DNA (dATP, dCTP, dGTP, dTTP) Primers : short (~ 25 nt), single stranded DNAs that tell the enzyme where to start DNA containing gene that you want to copy “template” PCR is carried out over a series of different temperatures: Thermal Cycling Denaturation: (95°C+) separates the double stranded template molecule Annealing: (~55°C) primers bind to single stranded DNA, tell enzyme where to replicate Elongation: (~72°C) enzyme adds dNTPs using the A:T, C:G rules Whole thing starts again, is an exponential process Thermus aquaticus: bacterium that the DNA polymerase is harvested from • Lives in hot springs • Enzymes don’t denature at hot temperatures (95°C+) • “ Taq DNA polymerase ” PCR amplifies a specific gene using an enzymatic reaction and the tendency of A to bind T and C to bind G DNA sequencing is a DNA replication process too, and involves adding labeled nucleotides that stop the replication reaction,...
View Full Document
This note was uploaded on 02/28/2008 for the course EBIO 1210 taught by Professor Demmig-adam during the Fall '07 term at Colorado.
- Fall '07