Team:USyd-Australia/Notebook

From 2014.igem.org

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  • Protocols
  • Primers
  • gBlocks
  • Notebook

    Date

    Day

    Activity

    Who?

    15/5 Thursday Preparation of common solutions:
    • LB
    • Stock Streptomycin (Sm) 200mg/mL, Chloramphenicol (Cm) 25 (mg/mL)
    • Sm200 and Cm25 plates
    Callum and Jeanne
    16/5 Friday Preparation of common solutions:
    • Stock Kanamycin (Km) 50 (mg/ml)
    • Km50 plates
    Abi and Tom
    22/5 Thursday Preparation of common solutions:
    • 2M NaOH
    • Stock Trimethoprim (Tm) 50mg/mL
    Restriction digest:
    • pK18 digested with HindIII, EcoRI, BamHI
    Agarose Gel Electrophoresis:
    • pK18 digests and undigested DNA
    • Post-stained with GelRed
    • Results...?
    Callum and Jeanne
    23/5 Friday Growing up gene source E. coli
    • JM109 containing pK18, for gene ____, on LB-Km
    • _____ containing R388, for gene ____, on LB-Tm
    • JM109 containing pUS44, for Pc-AttI1, on LB-Sm
    • Transform pUS41 by heat shock into Top10, grow on Sm (in hindsight, Top10 already has Sm resistance, so this was not valid)
    One Loopful of each culture from glycerol stocks was streaked onto appropriate selective media under aseptic conditions. All plates were incubated at 30 degrees over the weekend.
    Abi and Tom

    With thanks to: