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.
April 2, 2011
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.
April 21, 2011
April 23, 2011
April 27, 2011
April 28, 2011
Under Construction
May
May 18, 201
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.
May 20, 2011
Miniprep of P0440 in pSB1A2
Miniprep of P0440 in pSB1A2 was made following the Miniprep Protocol (Modified Bench Protocol: QIAprep Spin Miniprep Kit).
Assembly of Multiple Parts
Each of I13453, P0440, K331033, K331035, K331035, B0015, K249002, B0015, R0010 & B0034 were restricted where all odd parts were upstream and even parts were 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
May 26, 2011
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.
June
June 8, 2011
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).
June 9, 2011
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.
June 10, 2011
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.
June 11, 2011
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.
June 12, 2011
Overnight Cultures of K331033-K331035 & R0010-B0034
Overnight cultures of K331033-K331035 & R0010-B0034 grown following the Overnight Cultures Protocol.
June 13, 2011
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.
June 14, 2011
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).
Overnight Cultures of K331033-K331035 & R0010-B0034
Overnight cultures of K331033-K331035 & R0010-B0034 grown following the Overnight Cultures Protocol.
June 15-16, 2011
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.
June 22, 2011
PCR of pBAD-P0440 in pSB1K3
pBAD-P0440 was PCRed following the PCR Protocol.
June 23, 2011
Confirmation Gel of pBAD-P0440 (PCR)
pBAD-P0440 (PCR) was 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.
The gel ran at 100V for 90 minutes.
The gel was then stained in Ethidium Bromide for 10 minutes.
TABLE? PICTURE?
Overnight Cultures of pBAD-rbs-lumazine-dt & K331033-K331035
Overnight cultures of pBAD-rbs-lumazine-dt & K331033-K331035 grown following the Overnight Cultures Protocol.
June 24, 2011
Minipreps of pBAD-rbs-lumazine-dt & K331033-K331035
Minipreps of pBAD-rbs-lumazine-dt & K331033-K331035 were made following the Miniprep Protocol (Modified Bench Protocol: QIAprep Spin Miniprep Kit Using Microcentrifuge).
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