User:Elaine Marie Robbins/Notebook/Capstone/2012/06/20

MB Concentration Effects

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Methods MB concentration effects Mix 3 μL AuNP/TS primers, 1 μL dNTP, 2.5 μL PCR buffer, and 250 μL diluted cell extract containing 100,000 cells. Incubate at 30°C for 30 min. Add MB to each solution: 12.2 μL 100x diluted MB for MB/primers = 1 1.22 μL concentrated MB for MB/primers = 10 6.11 μL concentrated MB for MB/primers = 50 Incubate at 37°C for 60 min. Primers vs AuNP, incubation vs no incubation Make 4 solutions, each with 100,000 cells: Citrate AuNP/TS primer, incubated Mix 3 μL AuNP/TS primers, 1 μL dNTP, 2.5μL PCR buffer, and 250 μL diluted cell extract containing 100,000 cells. Incubate at 30°C for 30 min. Add 1.22 μL MB. Incubate at 37°C for 60 min. Citrate AuNP/TS primer, kept at room temperature Mix 3 μL AuNP/TS primers, 1 μL dNTP, 2.5μL PCR buffer, and 250 μL diluted cell extract containing 100,000 cells. Incubate at room temperature for 30 min. Add 1.22 μL MB. Incubate at 37°C for 60 min. Primers only, incubated Mix 6.5 μL 100x diluted primers, 1 μL dNTP, 2.5μL PCR buffer, and 250 μL diluted cell extract containing 100,000 cells. Incubate at 30°C for 30 min. Add 1.22 μL MB. Incubate at 37°C for 60 min. Primers only, kept at room temperature Mix 6.5 μL 100x diluted primers, 1 μL dNTP, 2.5μL PCR buffer, and 250 μL diluted cell extract containing 100,000 cells. Incubate at room temperature for 30 min. Add 1.22 μL MB. Incubate at 37°C for 60 min. Take fluorescence data for each solution. Data