Minnesota/21 July 2008

From 2008.igem.org

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1. Results from 07-18-2008 PROBLEM: Only terminator growth. RFP, MCherry, and TetR promoter had no growth. Make 2mL cultures of terminator in LB media - once have growth after 2 hrs with shaking @ 220rpm's, then make 5mL cultures. The 5mL cultures are composed of 4.5mL LB media and 0.5mL from each 2mL cultures.
2. Go to St. Paul: Get BioBrick parts from iGEM notebook - (1)E0030, YFP (2) J06504, RFP (3) R0040, TetR Promoter (4) I732077, GFP + LVA (5) E0032, YFP+LVA and (6) I732078, RBS+RFP+LVA+Term.
3. Transform parts from St. Paul: Biobrick parts from St. Paul stated above are to be transformed into TOP10 competent E. Coli cells. Incubate in 2mL cultures for 2 hours. Plate 200uL from cultures O/N in an incubator.
4. Order the following: (1) Dual promoter tailed primers to remove RBS, and (2) SSRA tag + Restriction sites tailed primers for GFP and YFP/RFP.
5. Biomedical Genomics Center: Contact them for sequencing problems to target the issue.