Contents |
Confirmation Gel of I1343, B0034, C0040 & B0015
- Each of I1343, B0034, C0040 & B0015 were restricted following the Restriction Protocol 4.
- A gel was made following the 1% Agarose Gel Protocol (100mL).
- The restrictions were loaded onto the gel with a 1kb ladder and 100bp ladder.
- The gel ran at 120V for 60 minutes.
- The gel was then stained in Ethidium Bromide for 10 minutes.
- Results: The parts were confirmed with the correct sizes.
Assembly of I1343 & B0034 and C0040 & B0015
- Each of I1343, B0034, C0040 & B0015 were restricted following the Restriction Protocol 1 where all odd parts are upstream and even parts are downstream.
- I1343 & B0034 were ligated together in pSB1A3 following the Ligation Protocol 1.
- C0040 & B0015 were ligated together in pSB1A3 following the Ligation Protocol 1.
- Both ligations were transformed following the Transformation Protocol 1.
Transformation of P0440 in pSB1A2 from 2010 Kit Plate
- 10µL of Milli-Q water and extraction of P0440 in pSB1A2 from the 2010 kit plate (10I) were mixed together.
- The transformation was done following the Transformation Protocol 1.
Miniprep of P0440 in pSB1A2
Miniprep of P0440 in pSB1A2 was made following the Miniprep Protocol (Modified Bench Protocol: QIAprep Spin Miniprep Kit Using Microcentrifuge).
Assembly of Multiple Parts
- Each of I13453, P0440, K331033, K331035, K331035, B0015, K249002, B0015, R0010 & B0034 were restricted following the Restriction Protocol 1 where all odd parts are upstream and even parts are downstream.
- I1343 & P0330 were ligated together in pSB1A3 following the Ligation Protocol 1.
- K331033 & K331035 were ligated together in pSB1A3 following the Ligation Protocol 1.
- K331035 & B0015 were ligated together in pSB1A3 following the Ligation Protocol 1.
- K249002 & B0015 were ligated together in pSB1A3 following the Ligation Protocol 1.
- R0010 & B0034 were ligated together in pSB1A3 following the Ligation Protocol 1.
- All ligations and a negative control (water) were transformed following the Transformation Protocol 1.
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