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From 2014.igem.org

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