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Team:UNIPV-Pavia/Notebook/Week1
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*Plate containing BBa_J23100 showed red colonies, as we expected: this part is inserted into plasmid BBa_J61002 which places the RFP downstream of the inserted part, which is a constitutive promoter. | *Plate containing BBa_J23100 showed red colonies, as we expected: this part is inserted into plasmid BBa_J61002 which places the RFP downstream of the inserted part, which is a constitutive promoter. | ||
Revision as of 18:00, 27 May 2008
 Home
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|
|---|---|---|---|---|
 Dry Lab
|  Wet Lab
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|  Protocols
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|
Notebook
| Week 1 | Week 2 |
|---|
Week 1: 05/19/08 - May 05/23/08
05/19/08
- Let’s start our IGEM 2008 experience! At first, we broke the punch tool…:)
- We used a scalpel to cut and resuspend the following 22 paper spots:
| BBa_I14032 | BBa_R0079 | BBa_R0062 | BBa_R0040 |
| BBa_R0082 | BBa_R0051 | BBa_J23100 | BBa_C0161 |
| BBa_C0062 | BBa_C0078 | BBa_C0179 | BBa_C0051 |
| BBa_C0012 | BBa_C0040 | BBa_I15010 | BBa_I15008 |
| BBa_I15009 | BBa_E0040 | BBa_E1010 | BBa_B0030 |
| BBa_B1006 | BBa_E0240 |
- We used tweezers to put the cut paper into tubes containing 10 μl of warmed TE buffer.
- We transformed 60 µl of TOP10 E. coli with 4 µl of DNA in TE for all 22 parts, plated transformed bacteria and incubated overnight at 37°C.
- We used LB medium previously prepared, with the suitable antibiotic added.
05/20/08
- After overnight incubation, the following 14 plates showed colonies:
| BBa_R0079 | BBa_R0062 | BBa_R0040 |
| BBa_R0082 | BBa_J23100 | BBa_C0062 |
| BBa_C0051 | BBa_C0012 | BBa_C0040 |
| BBa_E0040 | BBa_E1010 | BBa_B0030 |
| BBa_B1006 | BBa_E0240 |
- While the following plates did not:
| BBa_I14032 | BBa_R0051 | BBa_C0161 |
| BBa_C0078 | BBa_C0179 | BBa_I15008 |
| BBa_I15009 | BBa_I15010 |
- Plate containing BBa_J23100 showed red colonies, as we expected: this part is inserted into plasmid BBa_J61002 which places the RFP downstream of the inserted part, which is a constitutive promoter.
- We picked up one colony from every working plate to grow 5 ml cultures of transformed bacteria overnight.
- We re-transformed 60 µl of TOP10 with the remaining 6 µl of DNA in TE for BBa_I14032, BBa_R0051, BBa_I15008, BBa_I15009, BBa_I15010, BBa_C0161, BBa_C0078, BBa_C0179.
- We plated transformed bacteria and incubated them at 37°C overnight.
