Part 1: Preparation of Media
-Prepared luminous media, NaCl Agar, and seawater complete liquid medium.
-Prepared motility agar using tubes of mannitol and glucose, enriched with salt.
-Carbohydrate utilization tubes were prepared by adding salt to usual constituents of glucose and mannitol tubes.
 

Table 1.  Media Recipes

Luminous Media	NaCl Agar	Seawater Complete Liquid Medium
o    NaCl- 30.0g o    Agar- 20.0g o    NH4Cl- 5.0g o    Pancreatic digest of casein- 5.0g o    Yeast extract- 5.0g o    K2HPO4- 3.9g o    KH2PO4- 2.1g o    CaCO3- 1.0g o    MgSO4 7H2O- 1.0g o    KCl- 0.75g o    Tris buffer (1M solution pH 7.5)-50.0ml            Tris base- 0.3g            SDS- 1.0ml of 10% solution            Water to 50 ml o    Glycerol- 3.0ml	o    NaCl- 70.0g o    Distilled water- 1000.0ml o    Nutrient Agar- 15.0g            Beef extract- 3.0g            Peptone- 5.0g            Agar-agar- 15.0g            Distilled water to 1000ml	o    Pancreatic digest of casein- 5.0g o    Yeast extract- 3.0g o    Synthetic seawater- 750.0ml            NaCl- 27.0g            CaCl2- 0.3g            KCl- 0.6g            MgSO4 7H2O- 7.0g            Na-glycerophosphate- 0.2g            Tris buffer- 2.0g            Monosodium glutamate- 5.0g            Glucose- 1.0g            Vitamin B12- 1.0mcg o    Distilled water- 1.0L o    Glycerol- 3.0ml

Part 2: Incubation of Squid
-Obtained four squid from a Korean market.
-Incubated squid partially submerged in 3% salt solution at four degrees Celsius overnight.
-Observed periodically throughout the next day for presence of glowing colonies.

Part 3:  Isolation of Luminescent Bacteria
-Each squid was swabbed and samples were plated on luminous media and NaCl Agar.
-The following day, squid were checked again and new samples were swabbed and plated on luminous media and NaCl Agar.
-In addition, glowing colonies found on NaCl Agar were transferred to new luminous and NaCl plates.
-Each following day, plates were examined and then restreaked in order to obtain isolated colonies of the luminescent bacteria.

 Part 4:  Maintaining Growth
-Successfully isolated bacteria were transferred to seawater complete liquid media.
-Plates continued to be restreaked everyday in order to maintain growth.
 

Part 5:  Characterization of Luminescent Bacteria
-Gram stain, motility stab, wet mount, oxidase test, and carbohydrate utilization of mannitol and glucose were used.
 

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