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.
- Gel
Transformation of K325909 in pSB1C3 and K325229 in pSB1C3 from 2011 Kit Plate
- 10µL of Milli-Q water and extraction of K325909 in pSB1C3 from the 2011 kit plate (17m) were mixed together.
- 10µL of Milli-Q water and extraction of K325299 in pSB1C3 from the 2011 kit plate (17c) were mixed together.
- The transformation was done following the Transformation Protocol 1.
Minipreps of Multiple Parts
- Minipreps of K331022, K331008, K331024, xylE, K331024, K331007, K331022, K331008 & K331007 were done following the Miniprep Protocol (Modified Bench Protocol: QIAprep Spin Miniprep Kit Using Microcentrifuge).
Assembly of pBAD-rbs & lumazine-dt
- Each of pBAD-rbs & lumazine-dt were restricted following the Restriction Protocol 1
- pBAD-rbs & lumazine-dt were ligated together in pSB1C3 following the Ligation Protocol 1.
- The ligation was transformed following the Transformation Protocol 1.
Transformation of pBAD-P0440
- pBAD-P0440 in pSB1K3 was transformed following the Transformation Protocol 1.
Overnight Cultures of pBAD-rbs & lumazine-dt
- Overnight cultures of pBAD-rbs & lumazine-dt grown following the Overnight Cultures Protocol.
Assembly of K331033 & K331035 and R0010 & B0034
- Each of K331033, K331035, R0010 & B0034 were restricted following the Restriction Protocol 1 where all odd parts are upstream and even parts are downstream.
- K331033 & K331035 were ligated together in pSB1K3 following the Ligation Protocol 1.
- R0010 & B0034 were ligated together in pSB1K3 following the Ligation Protocol 1.
- Both ligations were transformed following the Transformation Protocol 1.
Overnight Cultures of K331033-K331035 & R0010-B0034
- Overnight cultures of K331033-K331035 & R0010-B0034 grown following the Overnight Cultures Protocol.
Assembly of Multiple Parts
- Each of K331033, K331035, R0010, B0034, pBAD-rbs & lumazine-dt were restricted following the Restriction Protocol 1 where all odd parts are upstream and even parts are downstream and with the pSB1K3 destination plasmid.
- K331033 & K331035 were ligated together in pSB1K3 following the Ligation Protocol 1.
- R0010 & B0034 were ligated together in pSB1K3 following the Ligation Protocol 1.
- pBAD-rbs & lumazine-dt were ligated together in pSB1K3 following the Ligation Protocol 1.
- All ligations were transformed following the Transformation Protocol 1.
Minipreps of R0010-B0034 & pBAD-rbs-lumazine-dt
- Minipreps of R0010-B0034 & pBAD-rbs-lumazine-dt were made following the Miniprep Protocol (Modified Bench Protocol: QIAprep Spin Miniprep Kit Using Microcentrifuge).
Overnight Cultures of K331033-K331035 & R0010-B0034
- Overnight cultures of K331033-K331035 & R0010-B0034 grown following the Overnight Cultures Protocol.
Assembly of K331033 & K331035 and R0010 & B0034
- Each of K331033, K331035, R0010 & B0034 were restricted following the Restriction Protocol 1 where all odd parts are upstream and even parts are downstream.
- K331033 & K331035 were ligated together in pSB1K3 following the Ligation Protocol 1.
- R0010 & B0034 were ligated together in pSB1K3 following the Ligation Protocol 1.
- Both ligations were transformed following the Transformation Protocol 1.
Confirmation Gel of pBAD-rbs-lumazine-dt & R0010-B0034
- Each of pBAD-rbs-lumazine-dt & R0010-B0034 were restricted following the Restriction Protocol 4.
- A gel was made following the 1% Agarose Gel Protocol (100mL), but 1.5% with 1.50g of Agarose and 100mL of 1x TAE.
- The restrictions were loaded onto the gel with a 1kb ladder and 100bp ladder.
- The gel ran at 100V for 60 minutes.
- The gel was then stained in Ethidium Bromide for 10 minutes.
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