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 Laboratory Fundamentals in Biological Engineering  posted by  duggu   on 12/8/2007  Add Courseware to favorites Add To Favorites  
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Abstract/Syllabus:

Engelward, Bevin, Drew Endy, Natalie Kuldell, Neal Lerner, Angela M. Belcher, and Atissa Banuazizi, 20.109 Laboratory Fundamentals in Biological Engineering, Spring 2006. (Massachusetts Institute of Technology: MIT OpenCourseWare), http://ocw.mit.edu (Accessed 07 Jul, 2010). License: Creative Commons BY-NC-SA

S. cerevisiae shown budding (top) and bound to CdS (bottom).

S. cerevisiae shown budding (top) and bound to CdS (bottom). (Image by Belcher lab.)

Course Highlights

This course features a complete set of labs and assignments.

» Learn more about OpenWetWare, an effort to promote the sharing of information, know-how, and wisdom among researchers and groups who are working in biology and biological engineering.

Course Description

This course introduces experimental biochemical and molecular techniques from a quantitative engineering perspective. Rigorous quantitative data collection, statistical analysis, and conceptual understanding of instrumentation design and application form the underpinnings of this course. The four discovery based modules include DNA Engineering, Protein Engineering, Systems Engineering, and Biomaterials Engineering. Additional information is available on the course Wiki (hosted on OpenWetWare.)

 

Syllabus

 

 

OpenWetWare logo.

OpenWetWare is an effort to promote the sharing of information, know-how, and wisdom among researchers and groups who are working in biology and biological engineering. OWW provides a place for labs, individuals, and groups to organize their own information and collaborate with others easily and efficiently. In the process, we hope that OWW will not only lead to greater collaboration between member groups, but also provide a useful information portal to our colleagues, and ultimately the rest of the world. 20.109 is hosted on OpenWetWare.

Welcome to 20.109!

For many of you this will be the first time in a research lab and for others it will not, but it is our goal to make this class a useful and fun introduction to experiments and techniques in biological engineering. There is not enough time to show you everything you'll need to know if you go on to do research, but after taking this class you should feel confident and familiar with some fundamental experimental approaches and lab protocols. You will develop good habits at the bench, ones that will increase the likelihood of success in your work and insure the health and safety of you and those around you. By the end of the semester, you should also be aware of good scientific practice, having had some experience with report writing, notebook keeping and publicly presenting your data. All of us involved in teaching 20.109 hope you will find it a satisfying challenge and an exciting experience that has lasting value.

Overview

This course introduces experimental biochemical and molecular techniques from a quantitative engineering perspective. Rigorous quantitative data collection, statistical analysis, and conceptual understanding of instrumentation design and application form the underpinnings of this course. There are four discovery based modules:

  1. DNA Engineering - design, construct, and use mammalian expression vectors to assess DNA recombination in mammalian cells.
  2. Protein Engineering - rationally modify an enzyme in vivo, then quantitatively assess its expression, purification profile, and reaction kinetics.
  3. Cell Engineering - use biologically-encoded devices to design and build a two color, bacterial photography system; characterize behavior of resulting system.
  4. Biomaterials Engineering - use yeast surface display to screen a peptide library for sequences with affinity for metals.

Layout of the Class

You will perform four series of experiments (called "modules") over the course of the semester. The modules differ in length and in the ways you will be assessed. The experiments and your assignments are:

 

MODULE #

TOPICS

ASSIGNMENTS

1

DNA Engineering

Written Report

2

Protein Engineering

Oral Presentation

3

Systems Engineering

Written Report

4

Bio-material Engineering

Oral Presentation and Written text

 

You will be working as pairs throughout the semester, but you will be submitting individual lab reports and giving individual oral presentations.

In addition to the assignments listed above there will be

  1. Daily Lab Quizzes - These are intended to refresh your memory about the experiment you are performing. They will not be hard and should take no more than 5 minutes at the beginning of lab.
  2. Homework Assignments - These will vary considerably. You may be asked to perform a calculation, draw a conclusion, and/or make a figure using the data you have collected.
  3. Laboratory Notebooks - You will record your data on the white pages of a bound notebook. The yellow, duplicate pages will be collected and evaluated by the TA.

Late work will be penalized 1/3 of a letter grade for each day late and will not be accepted after a week.

Grading

 

TOPICS

PERCENTAGES

DNA Engineering

25%

Protein Engineering

10%

Systems Engineering

25%

Bio-material Engineering

20%

Daily Lab Quizzes

5%

Homework Assignments

10%

Laboratory Notebooks

5%

 

Lab Attendance

Lab attendance is mandatory and there are no make-up labs. A family crisis or severe illness requiring attention from the infirmary and prohibiting you from all your coursework are acceptable reasons for missing lab and every effort will be made to accommodate you in these exceptional circumstances.

General Lab Policy

Things To Do

  1. Be on time. At the start of the lab period, there will be a short introduction to the experiment you will perform that day. It is unfair to your partner and to others in the lab if you are not up to speed when the work begins.
  2. Inform the instructor and/or TA if there is a problem. You will have their immediate attention if you have cut yourself (even if you consider it minor), if something broke and needs cleaning up, or if you are on fire.
  3. Be aware of all the safety devices. Even though the instructor and TA will take care of emergencies, you should know where to find the first aid kit, the chemical spill kit, the eye wash and the safety shower.
  4. Keep clutter to a minimum. There is a coat rack to hang your jackets and there are empty cabinets to store your backpacks. Anything left in the aisles is likely to be stepped on and is a hazard to everyone.
  5. Wash your hands before you leave the lab for the day.
  6. Be aware of others in the lab. Areas of the room may be crowded at times and you should take care not to disturb the experiments of others in the lab.
  7. Bring your lab notebook, lab manual and open mind to every lab meeting.

Things Not To Do

  1. Do not eat, drink, chew gum, smoke or apply cosmetics in the lab. Just being in lab makes your hands dirtier than you can imagine and you don't want to accidentally eat any reagent (see item 5 on "things to do" list).
  2. Do not put pieces of lab equipment in your mouth. It sounds obvious but you'd be surprised!
  3. Do not work with chemicals until you are sure of their safe handling. This includes some awareness of their flammability, reactivity, toxicity, and disposal.
  4. Do not use the phone or computer with gloves on your hands.

Calendar

 

 

NK = Dr. Natalie Kuldell

BE = Prof. Bevin Engelward

DE = Prof. Drew Endy

AB = Prof. Angela Belcher

 

SES #

LABS

LECTURERS

1

Orientation

NK

Module 1: DNA Engineering

2

DNA Engineering by PCR

BE

3

Clean and Cut DNA

BE

4

Agarose Gel Electrophoresis

BE

5

DNA Ligation and Bacterial Transformation

BE

6

Examine Candidate Clones

BE

7

Restriction Map, Article Discussion/Tissue Culture

BE

8

Lipofection

BE

9

FACS Analysis

BE

Module 2: Protein Engineering

10

Tools for Protein Engineering

NK

11

Assessing Beta-galactosidase

NK

12

Purifying Beta-galactosidase

NK

13

Student Presentations

NK

Module 3: Systems Engineering

14

Tools for Systems Engineering

DE

15

Basic Bacterial Photography: Black and White

DE

16

Advanced Bacterial Photography: 2 Color

DE

17

Device Characterization

DE

18

Measuring DNA, RNA, Protein

DE

19

RT-PCR Data Analysis

DE

Module 4: Bio-material Engineering

20

Screening Library

AB

21

Optimizing Panning

AB

22

Rescreening Gold Binders

AB

23

PCR of Gold Binding Candidates

AB

24

Student Presentations

AB

25

Analyze Sequence Data

AB




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