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Unknowns
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1st lab session Two unknown slants
One is gram+ and the other is gram -. Unknowns will be numbered Inoculate each unknown on to a Brain Heart Infusion Agar (BHIA) – subculture (working culture) 2. Inoculate each unknown on MacConkey Agar – do streaking for isolation Gram staining on each unknown Label the plates, slides, tubes with the appropriate unknown numbers. Incubate all at 37oC
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2nd lab session Examine MacConkey for growth No growth – gram positive
Growth – gram negative These results will confirm the gram staining results Do catalase test only on gram positive unknown Catalase + Staphylococcus, catalase – Streptococcus
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Gram negative unknown Examine the isolated colonies on MacConkey and determine the color of the colony. Pink – lactose fermenter White/colorless – lactose nonfermenter We do not need this information for gram positive.
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Non- fermenter TSI slant – only one Urea broth Fermenter
Tryptone broth – 1 MRVP – 2 Citrate slant – 1 Urea broth – 1 Motility - 1
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3rd lab session Match the results on your unknown with the results on the tables for the known bacteria.
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Table 3 Table 1 Table 2 Table 4
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