Ashu did a titer assay with her phage lysate. She started with 100µL of phage lysate (10) and did 7 1:10 dilutions down 107. She added 10ul of each dilution to 250µL of M. smeg and plated it on L-agar plates. Her 102 plate had 29 plaques. What is the titer of her lysate? O 290 pfu/ml.
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- You have conducted serial 10-fold dilutions and measured the cfu (colony forming units) of a Streptococcus pneumoniae culture and also the pfu (plaque forming units) of a phage (virus) that infects the bacteria. You counted 5 cfu in a 0.4 ml sample of a 106 dilution of the bacterial sample. You then counted 50 plaque-forming units (pfu) in a 0.25 ml sample of a 108 diluted sample of the phage culture. What are the cfu/ml of the S. pneumoniae and pfu/ml of the phage cultures before dilution? 5 x 106 cfu/ml and 2 x 109 pfu/ml 4 x 107 CFU/ml and 2 x 109 PFU/ml 1.25 x 108 cfu/ml and 10 x 1010 pfu/ml 1.25 x 107 cfu/ml and 2 x 1010 pfu/mlWhich of the following is a measure of CPE from a viral infection? 1. End point dilution assay 2. ELISA 3. Plaque Assay 4. Hemagglutination assay 5. 1 & 31- Why was the T4 and E.coli mixed together in top agar and then poured on the plate instead of just spreading the two together with a glass spreader on the plate? 2- When doing the titration and isolation of the T4 phage, E.coli diluted for you to 10e7 cells/ml. why was a diluted culture used? 3- After doing the titration of T4 phage, you find your dilution plates are all TNTC. how could this have happened? 4- Could you isolate phages that infect E.coli from raw sewerage? explain.
- A phage-infected bacterial culture was subjected to a series ofdilutions, and a plaque assay was performed in each case, withthe following results. What conclusion can be drawn in the caseof each dilution? Dilution Factor Assay Results(a) 104 All bacteria lysed(b) 106 14 plaques(c) 108 0 plaques•There are two figures- one for vector control plate & another PCRinsert+vector experimental plate. Control plate = 10 blue colonies , Ligation plate ( experimental) = 150 white , 8 blue. Interpret what each figure means, what is its importance (control and experimental)? Why do colonies grow on vector control plates? Why are both blue and white colonies in your experimental plate? What were the components of the agar plate & their use (AMP+Xgal+IPTG)? What do these suggest about amp sensitivity and resistance? Why did we use X-gal and IPTG? Can you think of any issue with your result? Is the recombination efficiency good? Is your transformation efficiency good? •Write a concluding sentence about this lab and the data you analyzed.A student completed a plate and liquid lysate followed by a plaque assay. The volume harvested for the plate lysate was 5.5mL and had a titre of 1.8x10^10 PFU/mL. The volume harvested for the liquid lysate was 9.5mL and had a titre of 1.1x10^10 PFU/mL. Which of these lysates had the highest viral yield? Liquid lysate had the highest yield Plate lysate had the highest yield They both have the same yield
- Which of the following is TRUE when one assay bacteriophage titers? You should: a) first mix the phages with a live bacterial culture and then pour the mixture on the agar plate b) directly add the phage dilution onto the surface of an agar plate c) add tryptic soy broth to the phage dilution and incubate overnight d) incubate a phage solution with live bacterial cells for several minutes. You must add soft agar to the mixture before pouring the content on the agar plateWrite a lab report on enumeration of lytic viruses : the plaque assay including the sections: introduction, Materials and Methods, Results and Discussion, and citing peer reviewed sources use this data for results section Group Number Number of Plaques Dilution Factor of Plate Original Concentration of Phage 1 58 10^3 5.80E+04 2 37 10^3 3.70E+04 3 44 10^3 4.40E+04 4 63 10^3 6.30E+04 5 36 10^2 3.60E+03 6 65 10^3 6.50E+04 7 260 10^2 2.60E+04 8 39 10^3 3.90E+04 9 178 10^2 1.78E+04 10 192 10^2 1.92E+04 11 274 10^2 2.74E+04 12 224 10^2 2.24E+04 Class Average 3.52E+04You are counting plaques on your plaque assay plates made from serial dilutions of your high titer lysate. Your 10-5 plate has 615 plaques although some are butting up against each other so it is difficult to get an accurate count. Your 10-6 plate has 42 plaques, and your 10-7 plate has only 1 plaque. Which plate would probably yield the most accurate titer calculation of your phage and why is it more trustworthy than the others?
- In a plaque assay, a phage has undergone _________ cycle when plaques are observed on the agar lawn post- incubation. What is the answer on the blank?Average plaques for bacteriophage A，B，C are 137，36，25. PFU/ml is average plaques multiply by the volume dilution multiply by the dilution factor. Show your working for each bacteriophage.Why is that bacterial slide agglutination technique important in diagnostic procedure?