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Standard Laboratory Program 10 The exercises in this program are designed to illustrate key principles of molecular genetics including genome organization, gene function, and DNA fingerprinting. Each exercise emphasizes an investigative approach where students draw conclusions from experimental results. Each exercise also illustrates fundamental biological concepts. The four exercises can be done independently or in sequence and are recommended for beginning and intermediate-level students.
1001. Anatomy and Evolution of the Genome 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. The exercise serves as a good introduction for determining the size of DNA molecules and provides an appreciation for the complexity of genomes from different organisms. 1002. Analysis of a Genome Segment 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.
Anatomy and Evolution of the Genome EXP-1001 1 2 3 4 5 6 7 8
DNA from plasmid pUC18 (lanes 1 & 5), lambda phage (lanes 2 & 6). calf thymus (lanes 3 & 7), and calf kidney (lanes 4 & 8) are separated by electrophoresis. The DNA samples for this exercise are supplied ready for electrophoresis.
Analysis of a Genome Segment EXP-1002
1 2 3 4 5 6 7 8
Lambda phage DNA (lanes 1, 2, 5, & 6) or a plasmid containing a segment of the phage genome (lanes 3, 4, 7, & 8) were digested with EcoR1 or EcoR1 + BamH1 prior to this electrophoretic separation.
1003. DNA Fingerprinting (An authentic analysis) DNA fingerprinting relies on the fact that DNA can be extremely variable in sequence from one individual to another and from one organism to another. Thus, the method has been used to identify individuals in forensic and paternity cases and to study genetic lineages of closely related organisms. These applications are discussed in the text and the latter application is illustrated in this experiment. In the exercise, students are given two DNA samples which are identified in the instructor guide. One sample is from sheep and the other from cow. In part A, the DNAs are digested with restriction enzymes and the sizes of the bands composed of highly repeated sequences are determined by comparison to standard DNAs of known lengths as shown below. In part B, the nonrepeated sequences are eliminated by a simple novel procedure. This step serves to enhance the banding patterns of the repeated sequences as shown in the gel on the right. From the results of this analysis, students determine which DNA sample is from cow and which is from sheep. This exercise requiring two 2-3 hour laboratory periods is the only authentic (non-simulated) DNA fingerprinting exercise available for the teaching laboratory.
DNA Fingerprinting EXP-1003 Part A Part B 1 2 3 4 5 6 7 8 1 2 3 4 5 6 7 8
DNA from cow (lanes 3 & 4) and sheep (lanes 6 & 7) were digested with EcoR1 or BamH1 prior to this electrophoretic analysis. DNA markers are shown in lanes 2 & 5. (Part A) Cow and sheep DNA are subjected to a simple novel procedure in order to eliminate nonrepeated sequences. The repeated sequences were then electrophoresed on this gel. (Part B)
1004. Genotype to Phenotype
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. In the second part of the exercise, the plasmids are introduced into E. coli by transformation and the color of the resulting colonies (blue or white) is then used to assess the functional status of the ß-galactosidase gene. This exercise can be completed within a single 3-hour laboratory session or two 2-hour laboratory periods. Contents of the Chemical Package (Click Here) Price List - Standard Laboratory Program 10
Individual Experiments Each of the individual experiments is supplied with the chemicals and laboratory guides needed for 16 students working in pairs. (Please see page 17 for additional details). If you chose one or more of the experiments below, you should also order Electrophoresis Package 3/4. Electrophoresis Package 3/4 provides sufficient agarose, gel stain and electrophoresis buffers for up to 6 of the individual experiments in this series. Electrophoresis Package 3/4 is also suitable for the experiments in Standard Laboratory Programs 3 and 4.
The following experiments include Chemicals and Instructions.
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