Registry/Measurement kit/Notebook/2007-6-18

• It appears that there was no growth on yesterday's transfer plates.

Plan for today

1. Bag those tet/chlor plates before they dry out (done)
2. Try the transformation again with the old ligation using the new plates (done)
3. Re-do the 3-way ligation, using different backbones (i.e., ones with tet, kan, chlor resistance, where applicable) (done)
4. Transform the above (done)
5. Check/update labels on the digest products (done)
6. start a culture of each of top10 and mg1655 (done)

3-way ligation

• see protocol

Promoter tester with RFP

• 2 ligation reactions:
  • B0032 (E/S) + J04650 (X/P) + 1AT3 (E/P)
  • B0032 (E/S) + J04650 (X/P) + 1AC3 (E/P)
• internally: Device A/2+4, labelled in green with relevant antibiotic resistance (T,C)

RBS tester with GFP

• 3 reactions:
  • R0040 (E/S) + I13401 (X/P) + 1AK3 (E/P)
  • R0040 (E/S) + I13401 (X/P) + 1AT3 (E/P)
  • R0040 (E/S) + I13401 (X/P) + 1AC3 (E/P)
• internally: Device B/1+3, labelled in black (K,T,C)

RBS tester with RFP

• 2 reactions:
  • R0040 (E/S) + J04650 (X/P) + 1AT3 (E/P)
  • R0040 (E/S) + J04650 (X/P) + 1AC3 (E/P)
• internally: Device C/1+4, labelled in blue (T,C)

Transformation

• Each reaction above was transformed into a culture of MG1655.

Cell culture

• Procedure:

1. Touch pipet tip to cell colony
2. Eject tip into tube with 5mL LB
3. Let grow overnight at 37°C

For tomorrow

• Check on MG1655/top10 cultures
• Check on both transformation attempts