BITC1402: BIOTECHNOLOGY METHODS - Week 4 – February 7, 9

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BITC1402:
BIOTECHNOLOGY METHODS




LOOMIS
-
PRICE



Continued next page

TENTATIVE CLASS SCHE
DULE


Assignments are in Seidman and Moore
, 2
nd

edition

(SM)
*
,
Seidman

(
Seid
), the on
-
line lab book, or as indicated.

LECTURE TOPIC

LECTURE
READING

LAB TOPIC

LAB
BOOK

Week 1


January
17, 19


1)
Introduction to a safe workplace.


2)
Working Safely in the Laboratory:

clothing,
gloves & eye protection



3)
Safety Video: Practicing Safe Science HHMI

4)
Glassware Washing Hazards.

5)
The Measurement of
Mass and
Volume


6) Introduction to Accuracy and Precision

7)
Math quiz, part 1

math bas
ics: exponents
and scientific notation
; the metric system


S
M
:

Ch 9
-
12;

533
-
537

Ch 13; 19,

20


Seid: 1, 3


Laboratory orientation: labware, equipment,



reagents, care and maintenance. Handling


chemicals

and solutions. Glass, paper biological waste
disposal.


Glassware cleaning.

Care and use of laboratory balances.


Use of volumetric glassware.


Using a Pasteur pipette
.


Using a Mohr pipette

Skills check:
10 mL Mohr pipet.


Skills check:
Use of the bala
nces








p 23

p 28


Week 2

January
24, 26


1)
Things you need to know before you start the
first lab:

2) Solution basics; making a solution at a given
concentration

3)
Math: solution/dilution, part 1

4
)
Water in the lab

5)
Theory of acids and bases

pH, pH meters and pH electrodes

6
)
Acids & Bases Problem Set

http://www.biology.arizona.edu/biochemistry/pr
oblem_sets/ph/ph.html






SM:

Ch 28 (I
-
III)

Ch 26

Ch 22


Seid:
2, 11,
15


Calibration and use of the pH meter and electrode.

Observe individual calibration of pH meter.

Lab exercise
:
Making up solutions of strong acids,
strong bases, weak acids and weak bases

Skills check:
Use of the pH meter


Demonstration

on the use of the program, Microsoft
Excel for making graphs and inserting them in a
Microsoft Word document.



Extended Learning Center:
Math & Science Computers

Using the directions on pages 9
-
11,
complete Lab
Assignment 1
.


P 35






BITC1402:
BIOTECHNOLOGY METHODS




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-
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Week 3


January 31, February 2


1
) Buffer basics

2) Mathematics Review: significant figures,
decimals, scientific notation.

3) Significant figures tutorial
s:

http://science.widener.edu/svb/tut
orial/sigfigures
.html

and
http://www.lon
-
capa.org/~mmp/applist/sigfig/sig.htm

)

4
)
Math Quiz: solution/dilution part 2:


Review dilutions



SM:

Ch 27


Seid 4, 12,
13


Lab Exercise:
Preparation of phosphate buffer stock
solutions.

29


Introduction to
Micropipettes.

Practice with instructor
observation.


Skills check:
micropipettors


Extended Learning Center:

Graphing, continued.

Lab Assignment 2




P
29



P 43

Week 4


February
7,

9


EXAM 1: Feb
9


1)
Spectrophotometry

2
) Before you start the third lab:

Basics of using a visible spectrophotometer

3
)
Mathematics review:
Descriptions of
d
ata:
p
opulations, samples, variables and
measurements of central tendency and dispersion
(mean,
variance, standard deviation, coefficient
of variation and normal distribution).






SM:

Ch 23, 25


Seid 16, 18,
19




Lab Exercise:
Micropipette Calibration Check.

Short write
-
up, with statistics


How to write up your data and
appropriate conclusions


Observe individual setup & use of spectrophotometer.



Lab Exercise:

Making a simple dye solution and visible


spectroscopy.



P
45







P 50

Week 5


February
14, 16


1) Protein purification overview
, part 1

2)
Mathematics: review

dilutions
;
applications
of Beer’s La
w


SM:

Ch 29, 33


Lab exercise:

Pipetting and Micropipetting check using
the spectrophotometer


First major

lab report

(
spectroscopy labs
)
.




P 61


BITC1402:
BIOTECHNOLOGY METHODS




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Week 6


February
2
1
, 2
3



1) Protein purification overview, part 2

2)
Mathematics: making a purification table;
math quiz: graphing using logarithmic data




SM:


Seid: 25


Finish up labs (catch
-
up day)




P 72

P 74

Week 7




February
28,
March 1


1) The Centrifuge.

2) The Centrifuge Video: Basic Centrifugation
Techniques.

3) Safety Video: Centrifugation Hazards HHMI

4)
Mathematics: Centrifuge calculations and
use of a nomogram; theory of dialysis



Molarity calculator

of Laboratory SolutionMolarity calculator



SM
:

CH 31, 32


Seid: 5



Overview of Protein Purification

Lab Exercise:

Preparation of saturated ammonium
sulfate

Lab Exercise:

Precipitation of rabbit serum


Lab Exercise:

Dialysis

Lab Exercise:

General assay methods;
Detection of
protein peak by UV
spectroscopy.


Writing a scientific hypothesis.

Writing methods and materials sections.



P 67

P 69


P70

P 72

Week 8


March
6, 8


Clean and Green: Research opportunities in
renewable energy.


Guest lecture from Dr. Kainer


Student presentations




Lab
Exercise:

Production of Biodiesel from vegetable
oils.


Lab Exercise:

Analysis of BD from vegetable oils






BD

Lab



March
13, 15



Spring Break






BITC1402:
BIOTECHNOLOGY METHODS




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Continued next page

Week 9


March
20, 22


EXAM 2: MARCH 22


1) Structure of Proteins
; primary structure
(amino acids)

and secondary
;
amino acid
problem set

2)
Mathematics:
Weight per
volume vs
Molarity,
Percents

solutions and
, Parts.

3
) Molarity calculator
http://chem.salve.edu/chemistry/molarity.htm
1)

4) Vide
o: Column Chromatography.


Handout


Lab Exercise:
. Setting up a DEAE
anion

exchange
cellulose column

Lab Exercise:

Purification of rabbit immunoglobulin by
ion exchange chromatography.


Short Lab Write
-
up.


P 74

Week 10


March
27, 29


1) Structure of
Proteins
; tertiary and quaternary
structure and function

2)
Mathematics: pH/pKa of amino acids and
zwitterions; the isoelectric point

3) Mathematic
s
: 3
-
D structure visualization and
manipulation

4)
Large Molecule Problem Set
http://www.biology.arizona.edu/biochemistry/pr
oblem_sets/large_molecules/large_molecules_pr
oblems.html


Handout


Continued.



Week 11

April 3, 5


1) Immunochemistry

2) Use of animals in the laboratory; production
of monoclonal and polyclonal sera

3)
Mathematics: serial dilutions; graphing
ELISA data







SM:

Ch 17, 24



Lab Exercise:
ELISA in the
identification of proteins.


Short

lab report





Hand
-
out

BITC1402:
BIOTECHNOLOGY METHODS




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-
PRICE



Continued next page

Week 12

April
10, 12


1) Upstream processing; preparation of cells and
fermentation


2)
Videos: (Fractionating a Cell
)


3) Mathematics: graphing fermentation yields



SM:

Ch 30


Seid: 17


Lab Exercise

Analysis of IgG fractio
ns

using gel
electrophoresis.


Second Major Lab Report

(Purification of IgG).



P 83

Week 13

April
17, 19


1)
Biotechnology and the Regulation of Medical
and Food Products

2) Enzyme kinetics; graphing enzyme
reactions



SM:

Ch 3, 4, 5


Lab Exercise:
Lysozyme: Enzymatic activity and
crystallization of lysozyme.


Short

lab report

(lysozyme).


P 93

Week 14

April
24, 26


EXAM 3: APRIL 22


1)
Introduction to Product Quality Systems

2)
Mathematics: dosage of
pharmaceuticals


SM:

Ch 6, 8


Seid: 9


Continue






Week 15

May 1, 3


1) Documentation : The Foundation of Quality

2) Quality Systems in the Laboratory


3) Safety Video, OSHA Safety Video, Safety in
the Research Laboratory (Radionuclide Hazards,
Chemical Hazards, Emergency Response).


SM:

Ch 7



Lab Exercise:

Production of a product under
GMP
Conditions.



Short
lab report (GMP)
.


Hand
-
outs


FINALS WEEK



May
8,
10


May
8
, REVIEW; May 1
0
,
FINAL EXAM