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Monday 8/30
- Use a different plate of L62-1 cells (HSP-LacI-GFP), pick 3 different colonies and re-perform the HSP-LacI-GFP
- Only use 0 mM Glucose and 5 mM Glucose
- Make new digests of L54 (HSP-LacI-RBS), E0430 (EYFP), and RFP-TetR backbone
- Ligate L54 to L56 (Lysis-Terminator), L54 to L57 (Another Lysis-Terminator), and L54 to E0430
- Transform aforementioned ligations into cells, along with L90 and L91 (both HSP-Lysis)
- Attempt PCR extraction and amplification of E. coli sigma 32 (HtpR)
Tuesday 8/31
- Fluorescence spectrometry shows that cells in 5 mM glucose produces significantly less GFP than those in no glucose growth medium.
- Transformations of L90-L95 grow slowly in 30 Celsius conditions; no colonies yet
- Ran gel electrophoresis of E. coli sigma 32 PCR product
- PCR product the right size! Order primers to turn sigma 32 (HtpR) into an RFC 10 BioBrick
- Completed first part in ligating L36 and L37 by PCR
Wednesday 9/1
- Colonies appeared on L91 and L95 plates
- Colony PCR of L91 and L95
- Ran gel electrophoresis of L91, L95 colony PCR products
- Completed second part in ligating L36 and L37 by PCR
- Digest, ligate, and transform PCR product of L36-L37 into cells
Thursday 9/2
Friday 9/3
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