Team:Panama/9 August 2010

August 9

High resolution agarose gel (for small fragments). For this we used two lanes (1 and 5) for the molecular weight. We used the Lamda DNA/Hind III marker from Biotools. We decided to run duplicates of the samples, so we have the Promoter and RBS on lanes 2 & 3; lane 4 is empty and once again, the Promoter and RBS are in lanes 6 & 7.

Ethydium Bromide staining:

The stock concentration was at 10mg/ml.

The concentration for gel staining was 0.5ug/ml.

We used 15ul of Ethydium Bromide in 300ml of dH2O. We stained it covered for 30 minutes. Afterwards, we washed it in water for 10 minutes each time.

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Digestion Repetitions:

1. Promoter (using iGEM restriction enzymes)

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2. Promoter (using INDICASAT restriction enzymes)

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3. RBS

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The reason behind testing different batches of the same restriction enzymes for the promoter was to find out the reason why we didn't see anything when we ran electroforesis gels. We had thought that maybe the restriction enzymes were not doing their job, so we tried digesting the parts with enzymes that INDICASAT provided.

PsB1C3 Plasmid transformation (Stock):

• LB plates with Cloranfenicol at a concentration of 33ug/ml.

• LB plates with Tetracycline at a concentration of 15ug/ml.