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). (Image by Belcher lab.)
Course Highlights
This course features a complete set of labs and assignments.
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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
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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:
- DNA Engineering - design, construct, and use mammalian expression vectors to assess DNA recombination in mammalian cells.
- Protein Engineering - rationally modify an enzyme in vivo, then quantitatively assess its expression, purification profile, and reaction kinetics.
- Cell Engineering - use biologically-encoded devices to design and build a two color, bacterial photography system; characterize behavior of resulting system.
- 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:
Layout of the class.
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
- 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.
- 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.
- 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
Grading criteria.
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
- 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.
- 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.
- 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.
- 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.
- Wash your hands before you leave the lab for the day.
- 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.
- Bring your lab notebook, lab manual and open mind to every lab meeting.
Things Not To Do
- 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).
- Do not put pieces of lab equipment in your mouth. It sounds obvious but you'd be surprised!
- Do not work with chemicals until you are sure of their safe handling. This includes some awareness of their flammability, reactivity, toxicity, and disposal.
- 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
Course calendar.
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
|
|
|