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Team:UNIPV-Pavia/Notebook/Week3Dec
From 2009.igem.org
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| + | ===Trasformation of Ligations=== | ||
| + | |||
| + | A5: BBa_Q0040 with BBa_I732017 (RBS-TETR-PTET-RBS-LACZ-TT)<br> | ||
| + | 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) <br> | ||
| + | |||
| + | Infect: | ||
| + | |||
| + | # X-GAL plate with 200 ul A5 | ||
| + | # X-GAL plate with 100 ul A5 | ||
| + | # Agar plate with 200 ul A6 | ||
| + | # 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. | ||
Revision as of 08:58, 20 June 2009
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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:
- X-GAL plate with 200 ul A5
- X-GAL plate with 100 ul A5
- Agar plate with 200 ul A6
- 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.
