By Jeffrey M. Becker
The targets of this moment version of Biotechnology: A Laboratory direction stay unchanged: to create a textual content that contains a chain of laboratory workouts that combine molecular biology with protein biochemistry options whereas supplying a continuum of experiments. The direction starts off with uncomplicated suggestions and culminates within the usage of formerly received technical event and experimental fabric. organisms, Sacchaomyces cerevisiae and Escherichia coli, a unmarried plasmid, and a unmarried enzyme are the experimental fabric, but the techniques and rules proven are broadly acceptable to different platforms. this article will function a superb relief within the institution or guide of introductory classes within the organic sciences.
Key positive aspects of this new edition:
* All routines and appendixes were updated
* comprises new workouts on
* Polymerase chain reaction
* Beta-Galactosidase detection in yeast colonies
* Western blotting
* New techniques brought for
* Large-scale plasmid isolation
* Yeast transformation
* DNA quantitation
* New appendixes extra, one in all which gives info on having access to organic details websites on the web (World broad Web)
* Use of non-radioactive fabrics and straightforward entry to microbial cultures
* Laboratory routines pupil proven for seven years
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Extra resources for Biotechnology A Laboratory Course
2. Use the quadrant streak method (described below) to prepare two streak plates (LB agar and YEPD agar) of the mixed culture. Repeat the procedure, using the continuous streak method (described below). Also streak (by both streaking methods) pure cul- Aseptic Technique and Establishing Pure Cultures 13 tures of E. coli onto LB agar and S. cerevisiae onto YEPD agar. Be sure to label each plate. 1) a. Draw quadrants on the outside bottom of an agar petri plate. bo Flame an inoculating loop and be sure to allow the loop to cool before introducing it into the broth culture.
To accomplish this task, (1) the work area is cleansed with an antiseptic to reduce the numbers of potential contaminants; (2) the transfer instruments are sterilized; for example, the transfer loop is sterilized by heating with a Bunsen burner before and after transferring; and (3) the work is accomplished quickly and efficiently to minimize the time of exposure during which the contamination of the culture or laboratory worker can occur. The typical steps for transferring a culture from one vessel to another are as follows: (1) flame the transfer loop; (2) open and flame the mouths of the culture tubes; (3) pick up some of the culture growth and transfer it to the fresh medium; (4) flame the mouths of the culture vessels and reseal them; and (5) reflame the inoculating Aseptic Technique and Establishing Pure Cultures 11 loop.
Weigh out 5 g of yeast extract, 10 g of peptone, and 10 g of glucose 20 Exercise 2 (dextrose) on a top-loading balance; dissolve these in distilled water by stirring with a magnetic stirrer. ) 2. Divide this broth solution into two equal parts by adding 250 ml each to two 500-ml Erlenmeyer flasks: Flask A: Make no further additions. Flask B: Add 5 g of agar. Swirl to disperse the agar. 3. Dispense the broth medium (Flask A) into 15 test tubes, adding 5 ml of broth to each tube. At this time, or during the autoclaving period, use of the pH meter may be demonstrated (see Exercise 2A in Appendix 1).
Biotechnology A Laboratory Course by Jeffrey M. Becker