Tag Archives: bacterial culture

Bacterial Cultures – Colonies Selected from Yesterday’s Colony PCRs

Inoculated 5mL of 1x LB + Kan50 (made by Steven on 3/23/11). Incubated O/N at 37C, 250RPM. Will perform mini preps tomorrow. The following samples were selected:

  • MM09 – #1, 2, 8
  • MM11 660bp – #1, 2, 8
  • MM10 2/8/11 – #1, 2
  • MM04 1/19/11 – #2, 3
  • MM11 3000bp – #3
  • MM04 1500bp – #4
  • MM06 1/19/11 – #1, 2
  • MM04 550bp – #1, 2
  • MM05 1/19/11 – #1, 2
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Bacterial Cultures – Colonies Selected by Steven from Steven’s Re-Streaked Plate

Inoculated 5mL of 1x LB + Kan50 (made by Steven on 3/23/11). Incubated O/N @ 37C, 250RPM. Will perform mini preps tomorrow. The following samples were selected (red text on the plate):

  • #9
  • #10
  • #13
  • #43
  • #45
  • #49
  • #56
  • #58
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Gigas Bacterial Challenge – 1hr & 3hr Challenges with Vibrio vulnificus

400mL O/N culture (1x LB+1% NaCl, 37C, 150RPM, 1L flask) of V.vulnificus (STRAIN??) and V.tubiashii (Strain: RE22) were pelleted (4300RPM, 25C, Sorvall ST-H750 rotor). Supe was removed and pellets were each resuspended in 50mL sea water. 1mL was taken from each to use for dilutions to determine colony forming units (CFU).

Two containers were set up with each containing 16 C.gigas, and air stone and 8L of sea water. The entire 50mL of V.vulnificus was added to one of the containers. 8 oysters were sampled (gill and mantle tissue) from each container at 1hr and 3hrs after the addition of V.vulnificus culture and immediately frozen on dry ice. Samples were stored @ -80C in the “Gigas Vibrio Exposure 1,3hrs 1/11/11″ box. Additionally, 1mL samples of the water were taken at each time to determine CFU in the water.

In addition to the samples taken above, the following tissues were taken from 5 control oysters at the 3hr time point and treated/stored in the same fashion as the others, specifically for assessment of cyclooxygenase tissue distribution analysis: muscle, digestive gland/gonad (difficult to differentiate)

All oysters were measured. Morphometric data is here.

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