| 1. Transformations: Transformations of genes/plasmids into cells and onto LB plates have grown.
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| 2. Primers: Transferring from glass tubes to 1.5mL tubes.
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| NOTE: Tube #12 looks darker than other primers possibly contaminated.
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| 3. Transformations continued:
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| a. Thaw cells on ice
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| b. Combine & incubate on ice (20 mins); 50uL of cells and 2uL plasmid.
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| c. Heat shock 1 minute @ 42C.
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| d. Incubate on ice for 5 minutes.
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| e. Recover in 250uL S.O.C. @ 37C for 2 hours
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| f. Plate 100uL and incubate O/N (overnight) @ 37C
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| *Refer to iGEM “transformation” section for details.
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| NOTE: only #6, #12, and #13 plasmids/genes grew on Dh5alpha competent cells. The other eleven plasmids used had to be re-done for a transformation onto TOP10 competent cells.
|
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