Phage Lab Essay

1598 WordsApr 11, 20127 Pages
Alisha Patel Genetics Lab TA: Vasant Chary March 21, 2012 Phage Titering of a Bacterial Culture and Recombination of Bacteriophage OBJECTIVES: The main objectives of this experiment included making dilutions of solutions, plating phage or bacteria, and determining the number of bacterial viruses or phage in a suspension. It was also conducted to demonstrate that two different mutants of phage T4 can exchange genetic material to give rise to wild-type phage. The experiment was used to distinguish mutants from wild-type by their host specificity. The recombination in bacteriophage was performed to determine the concentration of unadsorbed phage from the U series plates, total concentration from B series, and concentration of…show more content…
0-1 | 0 | 0 | 10-2 | 0 | 0 | 10-3 | 0 | 0 | B B 10-5 | 41, 47, 56, 44 (188) | 0 | 10-6 | 0 | 0 | 10-7 | 1.88 x 107 pfu/mL | 0 | Table 3: Number of Plaques on Each Tube Series Tube Series | Volume Plated (mL) | Dilution Factor | Bacteria Strain | Plaques | Titer | B | 0.1 | 10-3 | B | 484 | 4.84 x 106 | | 0.1 | 10-4 | B | 644; 384 avg: 514 | 5.14 x 109 | | 0.1 | 10-5 | B | 80 | 8.0 x 107 | | 0.2 | 10-3 | B | 670 | 3.35 x 106 | | 0.2 | 10-4 | B | 132; 604 avg: 368 | 1.84 x 109 | | 0.2 | 10-5 | B | 114 | 5.7 x 107 | K | 0.1 | 10 | K | 142; 660 avg: 401 | 4.01 x 103 | | 0.1 | 10-1 | K | 233 | 2.33 x 104 | | 0.1 | 10-2 | K | 63 | 6.3 x 104 | | 0.2 | 10 | K | 521; 924 avg: 723 | 3.62 x 102 | | 0.2 | 10-1 | K | 392 | 1.96 x 104 | | 0.2 | 10-2 | K | 19* | -- | U | 0.1 | 10-3 | B | 396; 464;113 avg: 324 | 3.24 x 106 | | 0.1 | 10-4 | B | 3* | -- | | 0.2 | 10-3 | B | 784; 404 594 | 2.97 x 106 | | 0.2 | 10-4 | B | 24* | -- | *Only count plates with 40-400 plaques/plate Table 4: Recombination Frequency Tube Series | Volume Plated (mL) | Dilution Factor | Plaques | Titer | B | 0.2 | 10-4 | 368 | 1.84 x 109 | K | 0.1 | 10-1 | 233 | 2.33 x 104 | U | 0.1 | 10-3 | 324 | 3.24 x 106 | Titer on Phage Stock: # of plaques/(volume plated x reciprocal of the dilution factor of the dilution tube used) Recombination Frequency= 2(titer on K)/(titer on B - titer on U) =2(2.33 x 104)/(1.84 x 109 - 3.24 x 106) =2.54
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