Week from June 15th, to June 21st, 2009

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June, 15th

• This week we wanted to complete the assembly of an aTc sensor and to test if lacZ works. Sequence analysis cannot be efficiently performed to I732017 or to A3 because of lacZ coding sequence length (>3 Kb), so we decided to check lacZ integrity assembling a constitutive promoter upstream of A3 and testing the ligation on X-Gal plates.

• Digestion

June, 16th

June, 17th

Trasformation of Ligations

A5: BBa_Q0040 with BBa_I732017 (RBS-TETR-PTET-RBS-LACZ-TT)
A6: BBa_J23100 with BBa_Q0040 (PCON-RBS-TETR-TT-PTET )

• 1 ul A5 in Top10 Competent
• 1 ul A6 in Top10 Competent

Mix 80 ul of SOC with 20 ul X-GAL (40 mg/ml) and infect LB+Amp Agar plates.

• Add 250 ul SOC in 50 ul A5+ Top10
• Add 250 ul SOC in 50 ul A6+ Top10

1 hr incubation at 37°C (falcons with shaking 220 rpm)

Infect:

1. X-GAL plate with 200 ul A5
2. X-GAL plate with 100 ul A5
3. Agar plate with 200 ul A6
4. X-GAL plate with 20 ul of BBa_E0240 from glycerol stock (negative control).

Expected results:

• Plates 1 and 2 should develop blue colonies
• Plate 4 should remain colorless

16 hr incubation at 37°C.

June, 18th

Experiment with Tecan F200

• Description
• Purpose:
• Materials & Methods
• Protocol
• Results

June, 19th

June, 20th

June, 21st