Team:Lethbridge/Notebook/Lab Work/Group1

From 2011.igem.org

(Difference between revisions)
Line 39: Line 39:
==April 1, 2011==
==April 1, 2011==
<i> Confirmation Gel of I1343, B0034, C0040 & B0015</i>
<i> Confirmation Gel of I1343, B0034, C0040 & B0015</i>
-
<br>- Each of I1343, B0034, C0040 & B0015 were restricted following the Restriction Protocol 4.
+
* Each of I1343, B0034, C0040 & B0015 were restricted following the Restriction Protocol 4.
-
<br>- A gel was made following the 1% Agarose Gel Protocol (100mL).
+
* A gel was made following the 1% Agarose Gel Protocol (100mL).
-
<br>- The restrictions were loaded onto the gel with a 1kb ladder and 100bp ladder.
+
* The restrictions were loaded onto the gel with a 1kb ladder and 100bp ladder.
-
<br>- The gel ran at 120V for 60 minutes.
+
* The gel ran at 120V for 60 minutes.
-
<br>- The gel was then stained in Ethidium Bromide for 10 minutes.
+
* The gel was then stained in Ethidium Bromide for 10 minutes.
-
<br>- Results: The parts were confirmed with the correct sizes.
+
* Results: The parts were confirmed with the correct sizes.
==April 2, 2011==
==April 2, 2011==
<i>Assembly of I1343 & B0034 and C0040 & B0015</i>
<i>Assembly of I1343 & B0034 and C0040 & B0015</i>
-
<br>- Each of I1343, B0034, C0040 & B0015 were restricted following the Restriction Protocol 1 where all odd parts are upstream and even parts are downstream.
+
* Each of I1343, B0034, C0040 & B0015 were restricted following the Restriction Protocol 1 where all odd parts are upstream and even parts are downstream.
-
<br>- I1343 & B0034 were ligated together in pSB1A3 following the Ligation Protocol 1.   
+
* I1343 & B0034 were ligated together in pSB1A3 following the Ligation Protocol 1.   
-
<br>- C0040 & B0015 were ligated together in pSB1A3 following the Ligation Protocol 1.  
+
* C0040 & B0015 were ligated together in pSB1A3 following the Ligation Protocol 1.  
-
<br>- Both ligations were transformed following the Transformation Protocol 1.
+
* Both ligations were transformed following the Transformation Protocol 1.
==April 21, 2011==
==April 21, 2011==
==April 23, 2011==
==April 23, 2011==
==April 27, 2011==
==April 27, 2011==
==April 28, 2011==
==April 28, 2011==
-
Under Construction
+
* Under Construction
Line 62: Line 62:
==May 18, 201==
==May 18, 201==
<i>Transformation of P0440 in pSB1A2 from 2010 Kit Plate</i>
<i>Transformation of P0440 in pSB1A2 from 2010 Kit Plate</i>
-
<br>- 10µL of Milli-Q water and extraction of P0440 in pSB1A2 from the 2010 kit plate (10I) were mixed together.
+
* 10µL of Milli-Q water and extraction of P0440 in pSB1A2 from the 2010 kit plate (10I) were mixed together.
-
<br>- The transformation was done following the Transformation Protocol 1.
+
* The transformation was done following the Transformation Protocol 1.
==May 20, 2011==
==May 20, 2011==
<i>Miniprep of P0440 in pSB1A2</i>
<i>Miniprep of P0440 in pSB1A2</i>
-
<br>Miniprep of P0440 in pSB1A2 was made following the Miniprep Protocol (Modified Bench Protocol: QIAprep Spin Miniprep Kit).
+
* Miniprep of P0440 in pSB1A2 was made following the Miniprep Protocol (Modified Bench Protocol: QIAprep Spin Miniprep Kit).
<br><i> Assembly of Multiple Parts</i>
<br><i> Assembly of Multiple Parts</i>
-
<br>- 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.
+
* 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.
-
<br>- I1343 & P0330 were ligated together in pSB1A3 following the Ligation Protocol 1.  
+
* I1343 & P0330 were ligated together in pSB1A3 following the Ligation Protocol 1.  
-
<br>- K331033 & K331035 were ligated together in pSB1A3 following the Ligation Protocol 1.
+
* K331033 & K331035 were ligated together in pSB1A3 following the Ligation Protocol 1.
-
<br>- K331035 & B0015 were ligated together in pSB1A3 following the Ligation Protocol 1.
+
* K331035 & B0015 were ligated together in pSB1A3 following the Ligation Protocol 1.
-
<br>- K249002 & B0015 were ligated together in pSB1A3 following the Ligation Protocol 1.
+
* K249002 & B0015 were ligated together in pSB1A3 following the Ligation Protocol 1.
-
<br>- R0010 & B0034 were ligated together in pSB1A3 following the Ligation Protocol 1.
+
* R0010 & B0034 were ligated together in pSB1A3 following the Ligation Protocol 1.
-
<br>- All ligations and a negative control (water) were transformed following the Transformation Protocol 1.
+
* All ligations and a negative control (water) were transformed following the Transformation Protocol 1.
-
<br>- Gel
+
* Gel
==May 26, 2011==
==May 26, 2011==
<i>Transformation of K325909 in pSB1C3 and K325229 in pSB1C3 from 2011 Kit Plate</i>
<i>Transformation of K325909 in pSB1C3 and K325229 in pSB1C3 from 2011 Kit Plate</i>
-
<br>- 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 K325909 in pSB1C3 from the 2011 kit plate (17m) were mixed together.
-
<br>- 10µL of Milli-Q water and extraction of K325299 in pSB1C3 from the 2011 kit plate (17c) were mixed together.
+
* 10µL of Milli-Q water and extraction of K325299 in pSB1C3 from the 2011 kit plate (17c) were mixed together.
-
<br>- The transformation was done following the Transformation Protocol 1.
+
* The transformation was done following the Transformation Protocol 1.
Line 89: Line 89:
==June 8, 2011==
==June 8, 2011==
<i>Minipreps of Multiple Parts</i>
<i>Minipreps of Multiple Parts</i>
-
<br>- 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).
+
* 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==
==June 9, 2011==
<i>Assembly of pBAD-rbs & lumazine-dt</i>
<i>Assembly of pBAD-rbs & lumazine-dt</i>
-
<br>- Each of pBAD-rbs & lumazine-dt were restricted following the Restriction Protocol 1
+
* Each of pBAD-rbs & lumazine-dt were restricted following the Restriction Protocol 1
-
<br>- pBAD-rbs & lumazine-dt were ligated together in pSB1C3 following the Ligation Protocol 1.  
+
* pBAD-rbs & lumazine-dt were ligated together in pSB1C3 following the Ligation Protocol 1.  
-
<br>- The ligation was transformed following the Transformation Protocol 1.
+
* The ligation was transformed following the Transformation Protocol 1.
   
   
==June 10, 2011==
==June 10, 2011==
<i>Transformation of pBAD-P0440</i>
<i>Transformation of pBAD-P0440</i>
-
<br>- pBAD-P0440 in pSB1K3 was transformed following the Transformation Protocol 1.
+
* pBAD-P0440 in pSB1K3 was transformed following the Transformation Protocol 1.
<br><i>Overnight Cultures of pBAD-rbs & lumazine-dt</i>
<br><i>Overnight Cultures of pBAD-rbs & lumazine-dt</i>
-
<br>- Overnight cultures of pBAD-rbs & lumazine-dt grown following the Overnight Cultures Protocol.
+
* Overnight cultures of pBAD-rbs & lumazine-dt grown following the Overnight Cultures Protocol.
==June 11, 2011==
==June 11, 2011==
<i>Assembly of K331033 & K331035 and R0010 & B0034</i>
<i>Assembly of K331033 & K331035 and R0010 & B0034</i>
-
<br>- Each of K331033, K331035, R0010 & B0034 were restricted following the Restriction Protocol 1 where all odd parts are upstream and even parts are downstream.
+
* Each of K331033, K331035, R0010 & B0034 were restricted following the Restriction Protocol 1 where all odd parts are upstream and even parts are downstream.
-
<br>- K331033 & K331035 were ligated together in pSB1K3 following the Ligation Protocol 1.  
+
* K331033 & K331035 were ligated together in pSB1K3 following the Ligation Protocol 1.  
-
<br>- R0010 & B0034 were ligated together in pSB1K3 following the Ligation Protocol 1.
+
* R0010 & B0034 were ligated together in pSB1K3 following the Ligation Protocol 1.
-
<br>- Both ligations were transformed following the Transformation Protocol 1.
+
* Both ligations were transformed following the Transformation Protocol 1.
==June 12, 2011==  
==June 12, 2011==  
<i>Overnight Cultures of K331033-K331035 & R0010-B0034</i>
<i>Overnight Cultures of K331033-K331035 & R0010-B0034</i>
-
<br>- Overnight cultures of K331033-K331035 & R0010-B0034 grown following the Overnight Cultures Protocol.
+
* Overnight cultures of K331033-K331035 & R0010-B0034 grown following the Overnight Cultures Protocol.
==June 13, 2011==
==June 13, 2011==
<i>Assembly of Multiple Parts</i>
<i>Assembly of Multiple Parts</i>
-
<br>- 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.  
+
* 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.  
-
<br>- K331033 & K331035 were ligated together in pSB1K3 following the Ligation Protocol 1.
+
* K331033 & K331035 were ligated together in pSB1K3 following the Ligation Protocol 1.
-
<br>- R0010 & B0034 were ligated together in pSB1K3 following the Ligation Protocol 1.
+
* R0010 & B0034 were ligated together in pSB1K3 following the Ligation Protocol 1.
-
<br>- pBAD-rbs & lumazine-dt were ligated together in pSB1K3 following the Ligation Protocol 1.
+
* pBAD-rbs & lumazine-dt were ligated together in pSB1K3 following the Ligation Protocol 1.
-
<br>- All ligations were transformed following the Transformation Protocol 1.
+
* All ligations were transformed following the Transformation Protocol 1.
==June 14, 2011==
==June 14, 2011==
<i>Minipreps of R0010-B0034 & pBAD-rbs-lumazine-dt</i>
<i>Minipreps of R0010-B0034 & pBAD-rbs-lumazine-dt</i>
-
<br>- Minipreps of R0010-B0034 & pBAD-rbs-lumazine-dt were made following the Miniprep Protocol (Modified Bench Protocol: QIAprep Spin Miniprep Kit).
+
* Minipreps of R0010-B0034 & pBAD-rbs-lumazine-dt were made following the Miniprep Protocol (Modified Bench Protocol: QIAprep Spin Miniprep Kit).
<br><i>Overnight Cultures of K331033-K331035 & R0010-B0034</i>
<br><i>Overnight Cultures of K331033-K331035 & R0010-B0034</i>
-
<br>- Overnight cultures of K331033-K331035 & R0010-B0034 grown following the Overnight Cultures Protocol.
+
* Overnight cultures of K331033-K331035 & R0010-B0034 grown following the Overnight Cultures Protocol.
==June 15, 2011==
==June 15, 2011==
<i>Assembly of K331033 & K331035 and R0010 & B0034</i>
<i>Assembly of K331033 & K331035 and R0010 & B0034</i>
-
<br>- Each of K331033, K331035, R0010 & B0034 were restricted following the Restriction Protocol 1 where all odd parts are upstream and even parts are downstream.  
+
* Each of K331033, K331035, R0010 & B0034 were restricted following the Restriction Protocol 1 where all odd parts are upstream and even parts are downstream.  
-
<br>- K331033 & K331035 were ligated together in pSB1K3 following the Ligation Protocol 1.
+
* K331033 & K331035 were ligated together in pSB1K3 following the Ligation Protocol 1.
-
<br>- R0010 & B0034 were ligated together in pSB1K3 following the Ligation Protocol 1.
+
* R0010 & B0034 were ligated together in pSB1K3 following the Ligation Protocol 1.
-
<br>- Both ligations were transformed following the Transformation Protocol 1.
+
* Both ligations were transformed following the Transformation Protocol 1.
<br><i>Confirmation Gel of pBAD-rbs-lumazine-dt & R0010-B0034</i>
<br><i>Confirmation Gel of pBAD-rbs-lumazine-dt & R0010-B0034</i>
-
<br>- Each of pBAD-rbs-lumazine-dt & R0010-B0034 were restricted following the Restriction Protocol 4.
+
* Each of pBAD-rbs-lumazine-dt & R0010-B0034 were restricted following the Restriction Protocol 4.
-
<br>- A gel was made following the 1% Agarose Gel Protocol (100mL), but 1.5% with 1.50g of Agarose and 100mL of 1x TAE.
+
* A gel was made following the 1% Agarose Gel Protocol (100mL), but 1.5% with 1.50g of Agarose and 100mL of 1x TAE.
-
<br>- The restrictions were loaded onto the gel with a 1kb ladder and 100bp ladder.
+
* The restrictions were loaded onto the gel with a 1kb ladder and 100bp ladder.
-
<br>- The gel ran at 100V for 60 minutes.
+
* The gel ran at 100V for 60 minutes.
-
<br>- The gel was then stained in Ethidium Bromide for 10 minutes.
+
* The gel was then stained in Ethidium Bromide for 10 minutes.
-
<br>- TABLE? PICTURE?
+
* TABLE? PICTURE?
==June 22, 2011==
==June 22, 2011==
<i>PCR of pBAD-P0440 in pSB1K3</i>
<i>PCR of pBAD-P0440 in pSB1K3</i>
-
<br>- pBAD-P0440 was PCRed following the PCR Protocol.
+
* pBAD-P0440 was PCRed following the PCR Protocol.
==June 23, 2011==
==June 23, 2011==
<i>Confirmation Gel of pBAD-P0440 (PCR)</i>
<i>Confirmation Gel of pBAD-P0440 (PCR)</i>
-
<br>- pBAD-P0440 (PCR) was restricted following the Restriction Protocol 4.
+
* pBAD-P0440 (PCR) was restricted following the Restriction Protocol 4.
-
<br>- A gel was made following the 1% Agarose Gel Protocol (100mL).
+
* A gel was made following the 1% Agarose Gel Protocol (100mL).
-
<br>- The restrictions were loaded onto the gel with a 1kb ladder.
+
* The restrictions were loaded onto the gel with a 1kb ladder.
-
<br>- The gel ran at 100V for 90 minutes.
+
* The gel ran at 100V for 90 minutes.
-
<br>- The gel was then stained in Ethidium Bromide for 10 minutes.
+
* The gel was then stained in Ethidium Bromide for 10 minutes.
-
<br>- TABLE? PICTURE?
+
* TABLE? PICTURE?
<i>Overnight Cultures of pBAD-rbs-lumazine-dt & K331033-K331035</i>
<i>Overnight Cultures of pBAD-rbs-lumazine-dt & K331033-K331035</i>
-
<br>- Overnight cultures of pBAD-rbs-lumazine-dt & K331033-K331035 grown following the Overnight Cultures Protocol.
+
* Overnight cultures of pBAD-rbs-lumazine-dt & K331033-K331035 grown following the Overnight Cultures Protocol.
==June 24, 2011==
==June 24, 2011==
<i>Minipreps of pBAD-rbs-lumazine-dt & K331033-K331035</i>
<i>Minipreps of pBAD-rbs-lumazine-dt & K331033-K331035</i>
-
<br>- Minipreps of pBAD-rbs-lumazine-dt & K331033-K331035 were made following the Miniprep Protocol (Modified Bench Protocol: QIAprep Spin Miniprep Kit Using Microcentrifuge).
+
* Minipreps of pBAD-rbs-lumazine-dt & K331033-K331035 were made following the Miniprep Protocol (Modified Bench Protocol: QIAprep Spin Miniprep Kit Using Microcentrifuge).

Revision as of 20:22, 15 September 2011


Contents

April

April 1, 2011

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.

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, 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.
  • TABLE? PICTURE?

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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