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An Introduction to Cell and Molecular Biology (LC5)

This Program is a complete laboratory course for teaching cell and 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 3 hour laboratory sessions. The course consists of 16 experiments carefully selected from 7 Laboratory Programs that are presented in a comprehensive integrated laboratory manuals. 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, a water bath, and a small centrifuge such as tabletop model or microcentrifuge. The price is about $52 per student per semester if these items are available in your teaching laboratory.

In this laboratory course, college freshman and sophomore-level students will be introduced to the molecular biology of the eukaryotic cell. In the first two sections of the course, students study topics in protein biology and biochemistry such as protein structure, function, molecular evolution, and the detection and molecular basis of human disease. Techniques used for these experiments include electrophoresis and the Western blot procedure. In the third section of the course, students localize enzymes in plant and animal cells and study the properties of a specific cell-surface receptor. Students perform a project of their own design in the fourth section of the course. The projects focus on the characterization of plant peroxidases. For the projects, assorted vegetables or plants are needed but not included. Experiments on the properties and structure of DNA are presented in the final section of the course. Here, students perform experiments that deal with genome organization, chromatin structure, and specific gene function. Techniques include DNA electrophoresis, cell fractionation, DNA isolation, restriction nuclease mapping, and basic cloning procedures.

Features

  • Suitability - Eukaryotic Cell and Molecular Biology for freshmen and sophomores.
  • Lab Schedule - One 3-hour lab session per week for a full semester.
  • Cost - $52 per student per semester excluding costs of manuals and equipment.

The experiments that comprise this course were selected from 7 Programs.

B1-1 Properties of DNA (View Individual Experiment)

The DNA molecule from a single human chromosome is about 4 cm long and the length of DNA in an individual is about 200 times the distance from the earth to the sun. Isolated DNA in a test tube is also a long, stiff molecule. When alcohol is added to a DNA solution, the DNA fibers precipitate and can be spooled onto a glass rod. This feature of DNA is illustrated in the exercise, which provides enough purified DNA for 16 students working in pairs to perform the experiment.

B1-2 Cell Fractionation and DNA Isolation (View Individual Experiment)

Students isolate nuclei from calf thymus tissue and examine them microscopically. The DNA is then extracted from the nuclei by a simple procedure that uses a detergent and alcohol. Microscopes and a small centrifuge are desirable but not absolutely necessary for the exercise.

B4-2 Extraction and Analysis of an Enzyme from Wheat . (View Individual Experiment)

Acid phosphatase is present in many plant tissues where it catalyzes the removal of phosphate groups from macromolecules at low pH. In this exercise, students prepare a cell-free extract from wheat germ and determine the amount of the enzyme present in the extract. The experiment offers practical experience with enzyme extraction procedures and is an excellent introduction to the analysis of enzyme activity and basic enzyme kinetics.

A colorimeter is desirable but not absolutely necessary for this exercise.

EXP-1001 1001. Anatomy and Evolution of the Genome (View Individual Experiment)

Common plasmids are simple DNA molecules which contain a few genes and regulatory elements. Most viral genomes are more complex. For example, the genome of phage lambda contains approximately 50 genes. About 4,000 genes are present in the E. coli genome while there is approximately 1,000 times more DNA in the genome of a mammal. This progression in genome complexity is the topic of this exercise. Here, students compare the electrophoretic patterns of restriction digests of a plasmid, phage lambda DNA, and cow DNA from thymus and kidney as shown in the figure below.

EXP-1002 1002. Analysis of a Genome Segment (View Individual Experiment)

Phage lambda DNA and a recombinant plasmid containing a segment of the phage lambda genome are provided as the starting point for this exercise. Students digest these DNAs with EcoR1 and BamH1 and then analyze the fragments as shown below. Determination of the size of the fragments enables the student to identify the precise region in the lambda genome that is contained within the plasmid.

EXP-1004 1004. Genotype to Phenotype (View Individual Experiment)

This exercise was designed to provide an exciting introduction to specific gene structure and function. In the experiment, students are given two plasmids (A and B) which are identified in the instructors guide. One plasmid (A) has a functional gene for the enzyme ß-galactosidase. The ß-galactosidase gene in the other plasmid (B) is inactive because it contains a segment of foreign DNA. In the first part of the exercise, students analyze restriction digests of both plasmids in order to determine which plasmid should have a functional ß-galactosidase gene.

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-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-801 801. Serum Proteins and the Western Press-Blot (View Individual Experiment)

Western blotting is one of the most powerful methods in molecular biology for identifying and characterizing specific proteins in complex protein mixtures. We have now streamlined western-blotting procedures so that the entire analysis can be performed during a single 3-hour, or two 2-hour laboratory sessions. In exercise 801, students use the procedure outlined below to identify albumin, transferrin and gamma globulins in serum and then to study the evolutionary relationships of albumin in vertebrates.

EXP-803 803. Tissue-Specific Isoenzymes in the Cow (View Individual Experiment)

Isoenzymes are different molecular forms of the same enzyme and five major lactate dehydrogenase (LDH) isoenzymes are found in vertebrate tissues. The amounts of the isoenzymes vary in a tissue specific manner and these differences can be readily detected by localizing LDH activity in an agarose gel after electrophoresis of tissue extracts. In this exercise, students prepare a tissue extract from calf thymus and then compare the LDH isoenzyme profile to those from calf serum, heart and muscle.

EXP-804 804. Peroxidase Isoenzymes in Corn (View Individual Experiment)

Peroxidase is associated with the plant cell wall and different forms of peroxidase are found in different plant tissues. These tissue-specific differences are revealed by this exercise where students examine the electrophoretic patterns of peroxidase in corn root and shoot extracts.

IND-2 Tissue Printing (View Individual Experiment)

Locating specific proteins and nucleic acid molecules in tissue sections is an important goal in cell biology. An effective and simple technique for this purpose is tissue printing which permits the localization of specific macromolecules in animal and plant tissues. Here students perform this technique to examine the tissue distribution of the enzyme peroxidase in plants. First, students section carrots, celery, and other vegetables with razor blades and transfer the proteins from the cut sections to nitrocellulose membranes by application of gentle pressure.

IND-5 Specificity of Albumin Binding. (View Individual Experiment)

The binding of an enzyme to its substrate is only one example of the many specific molecular interactions that occur in biological systems. An analogous binding process occurs with serum albumin which binds certain small molecular weight compounds and serves as a carrier molecule for these compounds in blood. In this exercise, students use an electrophoretic assay to examine the binding of various dyes to albumin. The results of this graphic analysis show that the binding of dyes to albumin is saturable, specific, compatible, and dependent on the native structure of the protein.

Prices

Catalog # Price Description
LC5 1152.51

The Chemical Package for 16 students working in pairs plus 17 student manuals and one instructor manual.
LC5-C $841.15

The Chemical Package for 16 students working in pairs plus one student manual and one instructor manual.
LC5-SM 20.57

Sample Student Manual (175 pages) plus one instructor manual.
LC5-X 178.47

Nine student manuals plus one Instructor manual.