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Cell Biology (LC2)

This Program is a complete laboratory course for teaching cell biology or introductory molecular biology at the college level. The program is designed for 16 students working in pairs and provides essentially all of the chemicals and instructions that are needed to teach sixteen 2-3 hour laboratory sessions. The course consists of 14 experiments carefully selected from Standard Programs 1, 2, 3, and 7 that are presented in a comprehensive integrated laboratory manual.

The Table of Contents of the manual and the catalog numbers of the experiments provided with the program are shown below. To use this program, you will need an Accessory Kit, the appropriate electrophoresis equipment, microscopes, and a water bath. A small centrifuge is required for 2 experiments. The price of the course is about $51 per student per semester if these items are available in your teaching laboratory.

In this laboratory course, your students will be introduced to the molecular biology of the eukaryotic cell. In the first section of the course, students study topics in protein biology and biochemistry such as protein structure, function, isolation, molecular evolution, and the detection and molecular basis of human disease. Techniques used for these experiments include electrophoresis (both native and denaturing), affinity chromatography, peptide mapping, and the Western blot procedure. In the second section of the course, students localize enzymes in plant and animal cells, perform cell fractionation procedures, and study the properties of a specific cell-surface receptor. Experiments on the properties and structure of DNA are presented in the final section of the course. Techniques include DNA electrophoresis and restriction nuclease mapping.

Features:

  • Suitability - Eukaryotic Cell Biology or Introductory Molecular Biology for sophomores and juniors.
  • Lab Schedule - One 2-3 hour lab session per week for a full semester.
  • Cost - $51 per student per semester excluding costs of manuals and equipment.

The experiments that comprise this course were selected from four Programs:

EXP-101 101. Electrophoretic Separation of Proteins (View Individual Experiment)

Students are introduced to the theory of separating proteins according to charge differences using electrophoresis. They then study four proteins and relate differences in their charges to their migration rates in an electric field. Each protein is a different color so that its progress during the separation can easily be followed.

EXP-102 102. Genetics and Sickle Cell Anemia (View Individual Experiment)

Many changes in the structure of hemoglobin have arisen by mutations. About one person in 100 carries a mutant hemoglobin gene, and these individuals have abnormal hemoglobin molecules in their blood. One of the most common abnormal hemoglobins is hemoglobin S, which causes sickle cell anemia. When the gene for hemoglobin S is inherited from both parents, all of the hemoglobin in the circulation is hemoglobin S and the individual suffers from severe anemia.

Exp-106 106. Protein Fingerprinting (View Individual Experiment)

A comparison of specific proteins from different species provides a powerful approach for establishing evolutionary relationships and for identifying organisms. A common approach used for this purpose is protein fingerprinting where electrophoretic properties of specific proteins are analyzed in different species. In this exercise, students use the approach to compare the forms of the enzyme lactate dehydrogenase that are found in serum of different mammals. Typical results of this graphic experiment are shown below.

EXP-201 201. Molecular Weight Determination (View Individual Experiment)

A first step in characterizing a protein often involves determining its molecular weight. From this information, different proteins may be compared and the number of amino acid residues in a protein can be determined. Here, students determine the molecular weight of two unknown proteins by comparing their electrophoretic migration with the migration of standard proteins. The protein standards and unknowns have been pre-stained so that your students can follow their progress during the separation as shown.

EXP-204 204. Peptide Mapping Analysis (View Individual Experiment)

SDS gel electrophoresis is used extensively to separate and identify denatured proteins. However, because this method relies on protein size alone, little information about proteins with the same molecular weight can be obtained. Peptide mapping is one of a number of techniques used to study the relatedness of similarly sized proteins. With this method, proteases are used to cut proteins into smaller peptide fragments and the fragments derived from two or more proteins are compared.

EXP-205 205. Protein Evolution and the Western Blot (View Individual Experiment)

The Western blotting procedure is rapidly replacing conventional methods for identifying and characterizing specific proteins in complex protein mixtures. This technique is used extensively for this purpose in the research laboratory and is increasingly being used in diagnostic medicine for detecting proteins of disease agents such as the structural core proteins of the AIDS virus. Here, students will perform this technique to examine the evolutionary distance between different mammals.

EXP-206 206. Affinity Chromatography (View Individual Experiment)

Purified proteins are often needed in the basic research laboratory and for diagnostic and therapeutic procedures. An effective technique for protein purification is affinity chromatography, which exploits a specific interaction between a protein and a complementary binding molecule. In this exercise, students isolate albumin from horse serum by affinity chromatography using a column matrix containing a reactive blue dye which binds specifically to the albumin molecule. They then use electrophoresis to analyze the isolated protein in order to verify the effectiveness of the procedure.

EXP-301 301. The Length of DNA Molecules (View Individual Experiment)

Electrophoresis in agarose gels is the most common method used for determining the size of DNA molecules. In this introductory exercise, students determine the length of an unknown DNA molecule by comparing its electrophoretic mobility with six DNA molecules of known size as shown.

EXP-302 302. Restriction Nuclease Mapping of DNA (View Individual Experiment)

Bacteriophage lambda is a DNA virus that attacks E. coli. Here, students dissect lambda DNA using the restriction endonucleases EcoR1 and BamH1 in order to identify specific sites, sequences, and structures along the phage genome. This single exercise enables students to explore a number of exciting topics in molecular biology, including the specificity of restriction endonucleases, DNA mapping strategies, complementary base-pairing of DNA, and the structure of a viral genome.

DNA STAINING PROCEDURE

EXP-303 303. Plasmid DNA Structure (View Individual Experiment)

Plasmids are small circular DNA molecules found in most bacteria. A plasmid can exist in different structural states and these states can be distinguished by their migration on agarose gels. In this experiment, students study these structures using enzymes and electrophoresis and show that the structures are interconvertible. This exercise provides an introduction to higher-order DNA structure which is thought to be important in the control of transcription and replication in bacteria and, perhaps, in higher organisms as well.

EXP-306 306. The Nucleosome Structure of Chromatin (View Individual Experiment)

The primary level of chromosome structure in eukaryotes occurs when the DNA molecule is wrapped around histone proteins into particles called nucleosomes. Evidence for this "beads on a string" model is derived from nuclease digestion studies. When nuclei are incubated with micrococcal nuclease, the enzyme cleaves the linker DNA between nucleosomes (the string) but not the nucleosomal core DNA (the beads).

EXP-701 701. Enzyme Cytochemistry (View Individual Experiment)

The concept that different enzymes are found in different tissues, cell types, and cell organelles is illustrated in this multipart exercise where students use contemporary techniques to localize specific enzymes in cells and tissues. Students are introduced to enzyme cytochemistry in an experiment on the germinating corn seed where they show that peroxidase is produced by the aleurone, a cell layer that surrounds the endosperm. They then characterize the subcellular distribution of peroxidase in giant onion epithelial cells and show that the enzyme resides in the cell wall.

EXP-702 702. Analysis of a Cell-Surface Receptor (View Individual Experiment)

Chemical signaling between cells in multicellular organisms is frequently mediated by cell-surface receptors. The receptors for neurotransmitters, protein hormones, growth factors, and plant lectins are a few of the many known examples of these important membrane components. In this exercise, students examine the cell location and properties of the receptor for the lectin concanavalin A. In the first experiment of the series, students use a concanavalin A-peroxidase complex in a microscopic assay to show that the specific receptor is found on the surface of their own cheek epithelial cells.

EXP-703 703. The Cell Nucleus (View Individual Experiment)

The exercise is introduced with a discussion of observations that were made in the early nineteenth century. Namely, that biological stains often show specific affinity for particular subcellular components. With this information, students use differential staining procedures to identify nuclear and cytoplasmic components in cells from thymus and onion root tip. In the second part of the exercise, cell nuclei are isolated from thymus tissue and their purity assessed by staining techniques.

Prices

Catalog # Price Description
LC2 1004.80

The Chemical Package for 16 students working in pairs plus 17 student manuals and one instructor manual.
LC2-C $824.77

The Chemical Package for 16 students working in pairs plus one student manual and one instructor manual (The student manual may be reproduced for educational purposes.)
LC2-SM 20.57

Sample Student Manual (148 pages) plus one instructor manual.
LC2-X 154.53

Nine student manuals plus one instructor manual.