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Category: Applied and Industrial Microbiology
Getting Started, Page 1 of 2
< Previous page | Next page > /docserver/preview/fulltext/10.1128/9781555818135/9781555811785_Chap01-1.gif /docserver/preview/fulltext/10.1128/9781555818135/9781555811785_Chap01-2.gifAbstract:
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.
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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.
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.
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.
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.
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.
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.
Commonly used laboratory equipment. From left to right: balance, pH meter, and microcentrifuge.
Commonly used laboratory equipment. From left to right: balance, pH meter, and microcentrifuge.
Commonly used laboratory equipment. (Left) incubator; (middle and right) water baths.
Commonly used laboratory equipment. (Left) incubator; (middle and right) water baths.
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.
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.
Schematic representation of a bacterial cell. (Reprinted from B. R. Glick and J. J. Pasternak, Molecular Biotechnology: Principles and Applications of Recombinant DNA, 2nd ed. [ASM Press, Washington, D.C., 1998], with permission.)
Schematic representation of a bacterial cell. (Reprinted from B. R. Glick and J. J. Pasternak, Molecular Biotechnology: Principles and Applications of Recombinant DNA, 2nd ed. [ASM Press, Washington, D.C., 1998], with permission.)
Bacterial population growth curve.
Bacterial population growth curve.
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.
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.
Aseptic transfer technique.
Aseptic transfer technique.
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, Molecular Biotechnology: Principles and Applications of Recombinant DNA, 2nd ed. [ASM Press, Washington, D.C., 1998], with permission.)
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, Molecular Biotechnology: Principles and Applications of Recombinant DNA, 2nd ed. [ASM Press, Washington, D.C., 1998], with permission.)
Representation of a eukaryotic animal cell. (Reprinted from B. R. Glick and J. J. Pasternak, Molecular Biotechnology: Principles and Applications of Recombinant DNA, 2nd ed. [ASM Press, Washington, D.C., 1998], with permission.)
Representation of a eukaryotic animal cell. (Reprinted from B. R. Glick and J. J. Pasternak, Molecular Biotechnology: Principles and Applications of Recombinant DNA, 2nd ed. [ASM Press, Washington, D.C., 1998], with permission.)
DNA spooled onto a glass stirring rod.
DNA spooled onto a glass stirring rod.
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.
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.
Molecular structure of the four DNA bases showing hydrogen bonding between thymidine and adenine (T and A) and guanine and cytosine (G and C).
Molecular structure of the four DNA bases showing hydrogen bonding between thymidine and adenine (T and A) and guanine and cytosine (G and C).