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Section 1 : Getting Started

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Getting Started, Page 1 of 2

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Abstract:

This section talks about documentation of laboratory experiments, use of equipment in the biotechnology laboratory, preparation of solutions, preparation and maintenance of microbial cultures, isolation of DNA by spooling, and investigation of the structure and bond strength of DNA. Microorganisms, especially bacteria, are very important to biotechnology. They can be used to replicate large quantities of DNA and to produce desired proteins. Microbes inhabit the environment all around us. Aseptic techniques are necessary to ensure that one is working only with desired microbes, not environmental contaminants. Bacterial growth media may be solidified with agar, a complex polysaccharide derived from algae. Most microorganisms cannot digest agar, making it an ideal solidifying agent. Agar solidifies at 40 to 42°C but does not melt until the temperature is raised to about 80 to 90°C. Solid media, in petri dishes, may be used to isolate individual bacterial clones necessary to establish pure cultures and recombinant DNA libraries. DNA is the genetic material that gives a person his or her inherited characteristics. The basic structure of DNA is identical in every organism. Ultraviolet (UV) spectroscopy can be used to analyze the structure of DNA. Acridine orange and a UV light source can be used to detect double- and single-stranded DNA. The series of exercises provided in the section includes topic such as documentation of laboratory, use of equipment, and investigating the structure and bond strength of DNA.

Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1

Key Concept Ranking

Chromosomal DNA
0.4751941
Sodium Dodecyl Sulfate
0.44967163
Baker's Yeast
0.40158558
0.4751941
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Figures

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Figure 1.1

These are pages from an older computation (comp) book. results are recorded in the beginning of the book with a reference to the calculations and preparations, which are recorded in the second half of the book. this method allows results to be easily read without obfuscation by complicated notes regarding the experimental setup.

Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Figure 1.2

These are pages from a newer comp book. formulas for defined algal media are shown, with calculations for specific quantities written on consecutive pages. see the error in calculations noted at the bottom of page 067.

Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Image of Figure 2.1
Figure 2.1

Equipment used in working with small volumes of reagents and cultures. Front, from left to right: microcentrifuge tube rack containing 1.5-ml microcentrifuge tubes, disposable tips for a 0.1- to 1 .0- μl micropipette, a 10-ml serological pipette inserted into a pipette aid, and sterile 15-ml conical centrifuge tubes. Rear, from left to right: bag to collect biohazardous waste, micropipette rack holding 100- to 1,000- μl and 0.1- to 10- μl digital micropipettes.

Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Image of Figure 2.2
Figure 2.2

Commonly used laboratory equipment. From left to right: balance, pH meter, and microcentrifuge.

Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Figure 2.3

Commonly used laboratory equipment. (Left) incubator; (middle and right) water baths.

Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Figure 3.1

Laboratory equipment used for preparing solutions. From left to right: collection bag for biohazardous waste, media bottles, test tubes in foam rack, Erlenmeyer flasks, graduated cylinders, and beakers.

Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Image of Figure 4.1
Figure 4.1

Schematic representation of a bacterial cell. (Reprinted from B. R. Glick and J. J. Pasternak, 2nd ed. [ASM Press, Washington, D.C., 1998], with permission.)

Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Image of Figure 4.2
Figure 4.2

Bacterial population growth curve.

Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Image of Figure 4.3
Figure 4.3

Streaking an agar plate to obtain isolated colonies (four-waystreak method). Pick up the inoculum using a flamed loop, and streak the first pass. For each subsequent pass, flame the loop before and after streaking. Overlap each pass only once or twice. Some researchers find it satisfactory to use a three-way method.

Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Figure 4.4

Aseptic transfer technique.

Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Image of Figure 5.1
Figure 5.1

A schematic “twisted ladder” representation of the double-helical DNA molecule. The rungs of the ladder represent the complementary base pairs, and the ladder standards represent the deoxyribose-phosphate backbone. (Reprinted from B. R. Glick and ). J . Pasternak, 2nd ed. [ASM Press, Washington, D.C., 1998], with permission.)

Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Image of Figure 5.2
Figure 5.2

Representation of a eukaryotic animal cell. (Reprinted from B. R. Glick and J. J. Pasternak, 2nd ed. [ASM Press, Washington, D.C., 1998], with permission.)

Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Image of Figure 5.3
Figure 5.3

DNA spooled onto a glass stirring rod.

Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Image of Figure 6.1
Figure 6.1

Partially denatured DNA. Double- stranded region shows G-C base pairs (three hydrogen bonds) and A-T base pairs (two hydrogen bonds). Single-stranded region shows loss of base-pairing; all hydrogen bonds are broken.

Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Image of Figure 6.2
Figure 6.2

Molecular structure of the four DNA bases showing hydrogen bonding between thymidine and adenine (T and A) and guanine and cytosine (G and C).

Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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References

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1. Kevles, D. J. 1998. The Baltimore Case: a Trial of Politics, Science, and Character. W. W. Norton & Co., New York, N.Y.
2. Wandersee, J. H.,, D. R. Wissing,, and C. T. Lange (ed.). 1996. Bioinstrumentation: Tools for Understanding Life. National Association of Biology Teachers, Reston, Va.
3. Reger, D. L.,, S. R. Goode,, and E. E. Mercer. 1997=. Chemistry: Principles & Practice, 2nd ed. Saunders College Publishing, Fort Worth, Tex.
4. Ausubel, F. M.,, R. Brent,, R. E. Kingston,, D. D. Moore,, J. G. Seidman,, J. A. Smith,, and K. Struhl (ed.). 1994. Current Protocols in Molecular Biology. Wiley Interscience, New York, N.Y.
5. Cappuccino, J. G.,, and N. Sherman. 1998. Microbiology: a Laboratory Manual, 5th ed. Benjamin/Cummings, Menlo Park, Calif.
6. Clark, D. P.,, and L. D. Russell. 1997. Molecular Biology Made Simple and Fun. Cache River Press, Vienna, III.
7. Rasmussen, A. M.,, and R. H. Matheson. 1990. A Sourcebook of Biotechnology Activities. National Association of Biology Teachers, Reston, Va., and North Carolina Biotechnology Center, Research Triangle Park, N.C.
8. Turner, P. C.,, A. G. McLennan,, A. D. Bates,, and M. R. H. White. 1997. Instant Notes in Molecular Biology. BIOS Scientific Publishers, Springer-Verlag, New York, N.Y.
9. Viehland, C. 1996. DNA spooling using baker's yeast. Stratagene Educational Materials Newsletter 1:2.
10. Watson, J. D. 1980. The Double Helix: A Personal Account of the Discovery of the Structure of DNA. W. W. Norton and Company, New York, N.Y.
11. Clerc, S.,, and Y. Barenholz. 1998. A quantitative model for using acridine orange as a transmembrane pH gradient probe. Anal. Biochem. 259:3029130295.
12. Darzynkiewicz, Z. 1990. Differential staining of DNA and RNA in intact cells and isolated cell nuclei with acridine orange. Methods Cell Biol. 33:285288.
13. Darzynkiewicz, Z.,, and J. Kapuscinski,. 1990. Acridine orange: a versatile probe of nucleic acids and other cell constituents, p. 291314. In M. R. Melamed,, T. Lindmo,, and M. L. Mendelsohn (ed.), Flow Cytometry Sorting, 2nd ed. John Wiley & Sons, New York, N.Y.
14. Freifelder, D. 1982. Physical Biochemistry. W. H. Freeman & Co., New York, N.Y.
15. Lehninger, A. L.,, D. L. Nelson,, and M. M. Cox. 1993. Principles of Biochemistry. Worth Publishers, New York, N.Y.
16. Watson, J. D.,, and F. H. C. Crick. 1953. A structure for deoxyribose nucleic acid. Nature 171:737738.
17. Watson, J. D.,, N. H. Hopkins,, J. W. Roberts,, J. A. Steitz,, and A. M. Weiner. 1987. Molecular Biology of the Gene, 4th ed. Benjamin/Cummings, Menlo Park, Calif.

Tables

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Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1
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Citation: Scheppler J, Cassin P, Gambier R. 2000. Getting Started, p 11-48. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch1

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