Agenda • REMINDER: Lab Report is due NEXT WEEK • 10 more Microworlds entries due in 2 weeks!! Will have time to work on it next week • Today, we may be doing lab 8 and 9a. Exercise 2: Transcription, Translation, Mutation Practice • Follow directions • Remember base-pairing rules: – DNA DNA – DNA RNA • Remember how to use this table Second base C U UUU U UUC UUA UUG Phe Leu CUU C A CUC CUA Leu UAU UCU UCC UCA Ser UAC CAU CGU CCC CCA Pro CAC CAA AAU AUC lle ACC AUA ACA GUA GUG U C CCU ACU Val UGC Cys UAG Stop AUU GUC UGU UCG CAG GUU Tyr UGA Stop A UGG Trp G CCG Met or start G UAA Stop CUG AUG G A Thr AAC AAA ACG AAG GCU GAU GCC GCA GCG Ala GAC GAA GAG His Gln Asn Lys Asp Glu CGU CGA U Arg G CGG AGU AGC AGA AGG C A Ser Arg U C A G U GGC C Gly GGA A GGG G GGU Part 3: Cell Cycle and Mitosis • Read background in lab carefully • We will go over this info in lecture Cell Cycle: • Interphase: G1, S, G2 • Cell may renter cell cycle or leave cell cycle (G0) G2 S Mitosis G1 Cytokinesis •DNA replicated in S •Nuclear division =Mitosis •Cell division =cytokinesis G0 MITOSIS ANIMATION Actual MITOSIS photography Part 3: Cell Cycle and Mitosis • CELL CYCLE = entire life of cell, including division – Interphase = not dividing – Mitosis = nuclear division – Cytokinesis = cell division Interphase (before Mitosis) • EVENTS: – G1: Cell grows and prepares to copy DNA – S: Cell copies its DNA – G2: Cell prepares to divide • APPEARANCE: – Chromatin (DNA) not condensed into chromosomes – WHY? – Nuclear envelope present MITOTIC PHASES • Mitosis – – – – Prophase Metaphase Anaphase Telophase • Cytokinesis Prophase EVENTS: 1. Chromatin condenses into chromosomes 2. Chromatids visible 3. Nuclear envelope dissolves 4. Centrioles (Spindle fibers) move to opposite poles APPEARANCE: • Nucleus looks blotchy (chromosomes) • Nuclear envelope dissolves Metaphase EVENTS: 1. Chromosomes line up in center of cell APPEARANCE: • • Dark chromosomes lined up in center of cell No nucleus Anaphase EVENTS: 1. Spindle fibers shorten 2. Chromosomes are pulled to opposite poles of cell APPEARANCE: • Two sets of chromosomes visibly separated from each other Telophase EVENTS: 1. Chromosomes unwind 2. New nuclear envelope reforms APPEARANCE: • Two dark areas of chromatin present at opposite ends Cytokinesis EVENTS: • Animals: Daughter cells pinch apart at cleavage furrow • Plants: Cell plate forms in center of cell APPEARANCE: • Two new (usually smaller) cells form Cell Division in Plants Plants differ from animals • Plant cells have no centrioles • Plant cells differ in Cytokinesis: – a cell wall forms between the dividing cells – called a cell plate Mitosis Review Part 3: Mitosis A. Modeling Mitosis with Playdoh B. Observing mitosis in plant cell slides – Practice you’ll do this on a quiz C. Estimating time spent in each phase Part 3: Mitosis C. Estimating time spent in each phase – – – – Identify the phase for 25 cells Get 3 other people’s data (100 cells total) Calculate the percent time spent in each phase (If you have 5 cells/100 in metaphase, then the cells spend ~5% of the time in metaphase.) Part 1: DNA Extraction • You can extract DNA from practically everything • Strawberries, peas, your lab partner’s brain! 1. Make lysis buffer • Add Water • Add Detergent – To extract DNA, we need to dissolve the membranes – Why not the cell wall? 1. Make lysis buffer • Water • Detergent • Add Salt – Causes proteins and carbohydrates to precipitate out of solution – We only want DNA 2. DNA extraction • Since your lab partner may object to your isolating brain DNA, we’ll use strawberries • Squash strawberry to increase the surface area exposed to detergent • It simply makes smaller pieces 2. DNA extraction • Add lysis buffer and continue squashing 2. DNA extraction • Filter mixture through filter paper • The DNA will be in the filtrate solution you capture 2. DNA extraction • Pour ice-cold alcohol (equal volume to strawberry filtrate) down side of tube to precipitate DNA • Keep it cold! 2. DNA extraction • DNA is is not soluble in alcohol and will precipitate out of solution • It will appear like slimy snot between the alcohol-water interphase Home DNA Extraction • http://www.pbs.org/wgbh/nova/teachers/acti vities/2809_genome.html • http://biology.about.com/c/ht/00/07/How_Ex tract_DNA_Human0962932481.htm