LAB PRACTICAL 2

LAB PRACTICAL 2 - LAB PRACTICAL 2 LAB 13 ISOLATING...

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LAB PRACTICAL 2 LAB 13: ISOLATING BACTERIOPHAGE WITH BACILLUS Virus Isolation: Phage Filtrate: Source: soil because bacillus lives in soil and phages that would infect bacillus would also be likely to be found in soil. 1.add some soil to EGB broth and vortex/centrifuge for 5 minutes 2. filter 1ml with .45u syringe filter Soft Agar Overlay: Why Soft agar: makes a confluent lawn of host cells because viral cannot grow with out the,. But soft enough so phages can distribute among others. 3. add .1ml of Bacillus culture and .1ml of soil suspension filtrate to tube of molten soft agar. 4. Pour into PCA plate and swirl to distribute. 5. When solidified, incubate overnight right side up. Plaque: 6. count the number of clear circles (plaques) 7. Titer= PFU/ml= (#plaques /Volume of Filtrate added) Host Specificity To enter a host cell, bacteriophages attach to specific receptors on the surface of bacteria, including lipopolysaccharides , teichoic acids , proteins or even flagella . This specificity means that a bacteriophage can only infect certain bacteria bearing receptors that they can bind to, which in turn determines the phage's host range. 1. Label 3 tubes: Control + Host+NonHost 2. Add No Bacterial, Host Bacterial, Other Bacteria 3. stab center of plaque and inoculate Host and NonHost tube 4. Compare tubes.
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LAB 15: STAPHLOCOCCUS Staph: NonMotile, Gram +, cocci, cat (+), facultative anaerobes
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This note was uploaded on 02/11/2011 for the course BIOMI 2900 at Cornell.

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LAB PRACTICAL 2 - LAB PRACTICAL 2 LAB 13 ISOLATING...

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