Richard Duong MCRO Lab 21 September 2010 Week 4 Pre-Lab Purpose: To examine and be able to quantitate the amount of bacteriophages replicating inside susceptible bacterial cells. We will be able to quantitate the amount of bacteriophage in an undiluted stock by the formula Original concentration of phage = (# of PFU/volume plated) x (1/dilution of the tube counted). Procedure: 1. Using the lab manual, correctly label the plates with the correct dilution factors before starting experiment. Label soft agar and broth as obtained from hot water bath. 2. A tube of original bacteriophage stock will be given out and using ten-fold dilutions, create 10^-1, 10^-2, and 10^-3 dilutions of the bacteriophage a. Flick bottom of phage tube with finger several times to mix, creating a “vortex effect” (very critical) that re-suspends virus without contaminating media or top of the tube. b. Pipettes will be sterile as long as they are inside their wrappers. Only the cotton – containing end will be exposed. Insert pumps gently onto the end of the pipette
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