TTT conference presentation ENGLISH

Presented by Rosa Meza-Acevedo

Exploring the Microbial World

http://www.123rf.com/photo_5320347_hand-holds-petri-dish-with-bacteria-culture.html

Center for Biophotonics, Science

and Technology (CBST)

National Science Foundation

UC Davis Mills College

Biophotonics

Definition: the study of the interaction of light with biological

material- where “light” includes all forms of radiant energy whose

quantum unit is the photon

Photonics• direct manipulation of

photons

• Illumination

• study of light energy

• Microscopic imaging

Bio• Life

Exploring the Microbial World

I. Sample microbial environment

II. Control microbes

III. Transform E. coli with Green Fluorescent

Protein (GFP)

Culture Sample Media

Media contain nutrients

Liquid and semisolid forms

Luria Broth (LB) Agar Petri plate

http://www.bestliveshopping.com/toys-165795011-B004MKHNJK-

Pre_made_LB_Agar_Plate_10_plates

How to make LB agar plates?

Materials

Erlenmeyer flask

Sodium chloride (NaCl)

Yeast extract

Tryptone

Agar powder

Deionized (DI) water

Sterile Petri plates

Tools for Sterilization LB Agar

http://sites.google.com/site/scienceprofonline/Lab-

gradeAutoclave2.JPG

http://www.sz-wholesale.com/shenzhen_China_products/Pressure-

Cooker_1.htm

http://www.missingfeatures.com/2007/06/25/building-

a-better-microwave/

Microwave Pressure cooker

Autoclave

Agar handling guidelines

Label bottom only with name, date and initials

(ex/ LB agar 11/4/11 RM)

When pouring, store agar plates right side up

When solidified, store in refrigerator up side down in

container

http://www.benchfly.com/video.php?video=164

Sampling Microbial Environment

Worksheet

Introduction

Hypothesis

Prediction (if/then)

Procedure

Materials

Methods

How to sample?

Sample collection is the most important step!

Unidentified organisms

Contamination

Swab replicates

II. Controlling Microbes

Controlling microbes worksheet

Introduction

Hypothesis

Prediction (if/then)

Procedure

Materials

Method

Procedure

Antibiotics

Ampicillin

Neomycin

Streptomycin

Tetracycline

Erythromycin

Penicillinhttp://www.debendiagnostics.co.uk/Antibiotic.html

Antiseptics

Iodine

Alcohol

Disinfectants

Sapolio

Clorox

http://www.bio.miami.edu/hare/culture.html

Learning Outcomes!

At the end of this session, students will have an

appreciation of &/or be able to:

Introduction to microbiology and bacteriology

Differentiate between fungi and bacteria growth

characteristics

Importance of antibiotic selection and resistance

genes

Impact of disinfectants and antiseptics in controlling

microbes

III. Transform E. coli with Green

Fluorescent Protein (GFP)

Transformation

Introduce foreign DNA

pGLO plasmid

GFP

Green Fluorescent Protein (GFP)

Jelly fish Aequoreavictoria

Visual marker

http://brainwindows.wordpress.com/2008/10/08/2008-nobel-prize-in-chemistry-to-gfp/

GFP changes genetic makeup

GFP changes genetic makeup

E. coli glowing coloniesE. coli colonies

http://askville.amazon.com/genetically-blue-eyed-parents-produce-brown-eyed-

child/AnswerViewer.do?requestId=2969837

Transform E. coli with Green

Fluorescent Protein (GFP)

Why teach?

Important modern biotechnology technique

Central framework of molecular biology

DNA RNA Protein trait/physical characteristic

Advantages

Compatible with 50 min class periods

Serves entire class of 32 students

Up to 4 students per group

Striking results!

pGLO Bacterial Transformation Kit