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Team:UNIPV-Pavia/Notebook/Week2Aug
From 2009.igem.org
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Week from August 10th, to August 16th, 2009
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August, 10th
- Digestion for:
| B1-13(E-S)(X2) | B2-5(E-S)(X2) | B3-5(E-X)(X2) |
| B4-2(E-X)(X2) |
- Gel run for all of them (only 1 ul for B1-13(E-S)(X2) in order to check the length)
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- Band purification for:
| B2-5(E-S)(X2) | B3-5(E-X)(X2) | B4-2(E-X)(X2) |
- Ethanol precipitation with sodium acetate for B1-13(E-S)(X2).
- Ligation:
- B5 = B1(E-S) + B4(E-X) in pSB1AK3
- B6 = B2(E-S) + B3(E-X) in pSB1AK3
- We incubated the ligation reactions at 16°C overnight.
August, 11st
- We transformed the overnight ligations of B5 and B6. We plated the transformed bacteria on LB agar plates + Kan and incubated the plates at 37°C overnight.
- Digestion for:
| BOL1(E-S) | R0010(E-X) | F2620MIT1(E-S) |
| K112808(E-X) |
- Gel run/cut/purification for all of them. All the bands were present at the right size!
- Ligation (20 ul final volume) for:
- A11 = BOL1(E-S) + R0011(E-X) in pSB1A2 (we call it A11 again, because the previous A11 had a deletion ad we threw it away)
- A15 = F2620MIT1(E-S) + K112808(E-X) in pSB1A2
- We incubated the ligations at 16°C overnight.
- M9 supplemented medium preparation (with glycerol). This first attempt showed precipitations...we will try tomorrow to dissolve MgSO4 and CaCL2 in ddH2O very slowly.
August, 12nd
August, 13rd
August, 14th
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