Awesome day in the lab today,
Made up a broth of 1g Tryptone and 0.5g Soduim chloride in 100ml sterilised water. this was then dispensed into universal jars (4ml per universal) and autoclaved at 121C for 15 minutes. Using aseptic techniques i the innoculated the broths with Kl terrigena, Kl pneumoniae and Kl oxytoca these were incubated over night at 30C. So hopefully they will have enough growth so i can add Kovacks reagent tomorrow (indole test) and i can hopefully differentiate the three culures. I have confirmed Listeria monocytogenes and so have made a broth and will hopefully make up stabs and slopes tomorrow. Alice has kindley donated a pure sample of A. baumanni, which i streaked onto a nutrient agar plate and also innoculated a nutrient broth and incubated at 30C overnight. I have also confirmed Streptomyces aureofaciens which i will broth culture tomorrow. Mike was amazing and explained loading dyes in detail, and how pH can alter substances, and how dyes can be used to range DNA banding during electrophresis (bromphenyl blue around 300bp). Then mike sprinkled a red powder (flourescens) into a beaker of water, because of the pH change the water began to glow like a yellow highlighter pen. Mike explained why this happened (outer shell valence and refraction of light) but i was happy with my glowing water xx Next Nicola explained how she was to cut out DNA from the agarose gel she had just ran and the purpose of this. Then i watched as she cut out each band of target DNA so that tomorrow the agarose gel can be dissolved away to leave behind the desired DNA. I Iooked at the kit to do this and it is in no way as simple as it seems. Even had time for an ethics debate with Dan, Sophie, Rosie and Kamilla.
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