5-1 Experiment 5 ELECTROPHORESIS OF PROTEINS IN POLYACRYLAMIDE GELS Introduction Proteins posses charged amino acid side chains on their surfaces. This creates a pH dependent net charge of proteins. The net charge determines many a proteins physical prope
1-1 Experiment 1 WEAK ACID BUFFERS Introduction A large fraction of the constituents of cells are weak acids, and some are weak bases; for example: proteins and individual amino acids, nucleic acids and individual nucleotides, fatty acids, and most metabo
4-1 Experiment 4 PREPARATION OF AN ENZYME Introduction Life science research is an integrated enterprise connecting molecular, biochemical, cellular, developmental, genetic and evolutionary biology. These disciplines have generated enormous amounts of inf
2-1 Experiment 2 SPECTROPHOTOMETRIC METHODS: PROTEIN DETERMINATION Introduction Chemical analyses are part of almost every investigation in biochemistry. The substances to be measured are present in milligram, microgram, or even nanogram amounts, which is
Practice Problems: PCR Primer Design
1. Calculate the volumes of the ingredients to prepare a PCR reaction mix. The final volume of the reaction mix is 50
l, which includes 5 l of the target DNA (target DNA is approximately 1.0 ng to 0.4 ng per reaction).
Freeze-dried strawberry powder improves lipid profile and lipid
peroxidation in women with metabolic syndrome: baseline and post
Arpita Basu*1, Marci Wilkinson1, Kavitha Penugonda1
Quiz 1: Lab Period 3 (April 9, 2013)
Name: _KEY_ Room_
After 0.2 ml of 0.2 mM pH indicator dye Y was mixed with 0.8 ml of buffers of pH
values at 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, and 8.5, absorbance values at 600 nm
were determined to be 0.80, 0
Repair of a dsDNA break!
This process of strand invasion or
strand exchange requires specialized
factors that help the ssDNA region
nd the homologous dsDNA. !
The broken chromosome 3 end(s) are used as primers for synthesis of DNA
missing from the break.
Spontaneously generated study questions (currently lacking solutions).
Given 200 mL of 0.1 M Phosphate buffer pH = 6.5, and 25 mL of 1 M HCL, what is the final pH?
Given 300 mL of 0.05 M Tris buffer pH = 7.8, and
Using the Spectrophotometer
In this exercise, you will learn the basic principals of spectrophotometry and and serial dilution
and their practical application. You will need these skills to complete other exercises throughout
the semester. It
Zn Acetate staining and the Coomassie Blue staining of the GAF-4 purification scheme.
Zn Acetate Stain
Coomassie Blue Stain
20 l Post-DNase I Extract
5 l Post-DNase I Extract
20 l Chitin Affinity
Answers to Practice Problems: PCR Primer Design
1. The reaction mix contains the following reagents (standard concentrations) per reaction:
34.5 l H2O
5 l 10X PCR buffer solution
(1X PCR buffer solution: 10 mM TrisHCl pH 9.0, 50 mM KCl, and 1.5 mM MgCl2)
Experiment 7 (cont.)
pDNA Prep. Day 1
Mode of Action
Mechanism of Resistance
Derivative of penicillin. Interferes with cell
cleaves amp, destroying it.
Practice Problems: Experiment 1
Consult as needed: Table 1.1 pKa values for buffers in dilute solution
1.1. What volume of glacial acetic acid (17.6 N) and weight of sodium acetate3 H2 O (F.W. = 136) is
required to make 100 ml of 0.20 M buffer, pH 4.06? (
0INTRODUCTION TO THE LABORATORY Each student is to perform the exercises individually in any order. 1. Reproducible pipeting Pipetman single sample pipettes 2. Spectrophotometry: Theory and Operation Schimadzu Spectrophotometer (Photometric and Spectrum f