Plaque assay for detecting lysogeny this is a qualitative screening of e coli strains for lysogeny the basis of the w3104 is a known bacteriophage lambda lysogen, and will play the role of positive control in the assay mg1655 is the negative control. Interactive laboratory microbiology basic, applied & health sciences : v microbial physiology 1-8 : physiology 1 oxygen requirements objectives/key words oxygen requirements of microbes aerobes anaerobes bacteriophage titer no mouth pipetting. This assay is the most widely used technique for the isolation of virus and its purification, and to optimize the viral titers the basis of plaque assay is to measure the ability of a single infectious virus to form a “plaque” on a concurrent monolayer culture cells. Titer) of bacteriophage t2 to perfor m a plaque assay, a sample to be tested is first serially diluted to perfor m a plaque assay, a sample to be tested is first serially diluted agar overlays are then inoculated with the virus and a host bacterium and poured into petri plates. Figure 1: basic structure of bacteriophage (madigan and martinko, 2006) enteric bacteria are normal inhabitants of the intestines of humans and other animals (davis, 2005) but are often isolated from aquatic ecosystems after sewage has been introduced into the environment.
The purpose of plaque assays using serially-diluted bacteriophage is typically done to calculate the titer of your bacteriophage sample “titer” refers to the number of infectious virions (aka plaque-forming units or pfu) per volume of viral stock. Determination of a bacteriophage titer for all of the tests performed in lab, be able to identify: the media used, the chemical being tested, a positive and negative result, the microbial enzyme(s) involved, and products being produced. Sauer lab home in some cases, the titer of the phage is too high relative to the amount of bacteria in the 1:1 mix used for the transduction infection you are aiming for a multiplicity of infection (moi) of 05-1 ref is the earliest report i know of bacteriophage-dependent transduction in e coli this report is notable because it.
A major limitation with traditional phage preparations is the variability in titer, salts, and bacterial contaminants between successive propagations here we introduce the phage on tap (pot) protocol for the quick and efficient preparation of homogenous bacteriophage (phage) stocks this method produces homogenous, laboratory-scale, high titer (up to 1010-11 pfu∙ml-1), endotoxin reduced. A reliable method for storage of tailed phages only ten out of nineteen phages from nine families tested survived over 10 years without a loss of phage titer, the common e coli laboratory strain c600 appeared to be the only poor host for phage storage. The reuse or reproduction of any of the information, design or layout contained in this web site without the permission of antibody design labs is prohibited comments are closed company information.
Lab 5: bacteriophage 5 february 2013 introduction: a bacteriophage is essentially a virus that specifically infects bacteria a bacterial cell infection progresses in much the same way as a eukaryotic cell. The number of lytic plaques on the “1:1,000,000” plate was counted and the titer of the bacteriophage was calculated using the formula: number of lytic plaques dilution factor 18 = number of bacteriophages per ml of viral sample 5 results a) bacteriophage effect on escherichia coli population growth in table 1, a higher absorbance was. Bacteriophage are among the most diverse and numerous microbes inhabiting our planet yet many laboratory activities fail to engage students in meaningful exploration of their diversity, unique characteristics, and abundance. Start studying exp 33 - bacteriophage titration learn vocabulary, terms, and more with flashcards, games, and other study tools.
702/10702 spring 2005 question 1 (continued) to set up an experiment, you mix 3 ml of undiluted pnk/kbs1 cells (titer of 4 x 1011 cells/l) with 2 ml of p1 phage with a titer of 108 pfu/ml d) determine the moi of this experiment. The viral titer is a quantitative measurement of the biological activity of your virus and is expressed as plaque forming units (pfu) per ml to calculate the viral titer, take your plates out of the incubator and examine them. Phages are available year round 150 µl of phage will arrive in a plastic vial at titer of 1 x 107 the the concentration of infective viral particles per milliliter of culture suspension is referred to as the titer.
Lab 1: phage titration phage titration (determination of the number of phage particles in a stock) is an important molecular biology technique when genetic libraries in phage vectors are screened for positive clones, the plates that are being screened should have approximately 50-500 phage plaques per plate for optimal results. Exam on monday july8th, 2013 learn with flashcards, games, and more — for free. Determine the concentration of bacteriophage in a solution with serial dilution and the plate-pour method.