Engineering BioBrick vectors from BioBrick parts/Colony PCR - Revision history

Vaishnavi Ananth at 09:39, 19 November 2009

2009-11-19T09:39:21Z

←Older revision		Revision as of 09:39, 19 November 2009
Line 42:		Line 42:
	#Image gel with 302 nm transilluminating ultraviolet light using an ethidium bromide camera filter and an exposure time of 614 milliseconds.		#Image gel with 302 nm transilluminating ultraviolet light using an ethidium bromide camera filter and an exposure time of 614 milliseconds.

-	==~~BioStream~~ version==	+	==BioCoder version==
-	Following is ~~theEngineering~~ BioBrick vectors from BioBrick parts/Colony PCR protocol in ~~BioStream~~, a high-level programming language for expressing biology protocols. What you see here is the auto-generated text ouput of the protocol that was coded up in ~~BioStream~~ (see Source code). More information about ~~BioStream~~ can be found on my home page. Feel free to mail me your comments/ suggestions.[[User:Vaishnavi Ananth\|Vaishnavi]]	+	Following is the Engineering BioBrick vectors from BioBrick parts/Colony PCR protocol in BioCoder, a high-level programming language for expressing biology protocols. What you see here is the auto-generated text ouput of the protocol that was coded up in BioCoder (see Source code). More information about BioCoder can be found on my home page. Feel free to mail me your comments/ suggestions.[[User:Vaishnavi Ananth\|Vaishnavi]]
	====Text Output====		====Text Output====
	[[Engineering BioBrick vectors from BioBrick parts/Colony PCR protocol]]		[[Engineering BioBrick vectors from BioBrick parts/Colony PCR protocol]]

Vaishnavi Ananth at 06:23, 1 October 2009

2009-10-01T06:23:12Z

←Older revision		Revision as of 06:23, 1 October 2009
Line 41:		Line 41:
	#Also electrophorese 1 μg of 2-log DNA ladder to verify the length of each PCR product.		#Also electrophorese 1 μg of 2-log DNA ladder to verify the length of each PCR product.
	#Image gel with 302 nm transilluminating ultraviolet light using an ethidium bromide camera filter and an exposure time of 614 milliseconds.		#Image gel with 302 nm transilluminating ultraviolet light using an ethidium bromide camera filter and an exposure time of 614 milliseconds.
		+
		+	==BioStream version==
		+	Following is theEngineering BioBrick vectors from BioBrick parts/Colony PCR protocol in BioStream, a high-level programming language for expressing biology protocols. What you see here is the auto-generated text ouput of the protocol that was coded up in BioStream (see Source code). More information about BioStream can be found on my home page. Feel free to mail me your comments/ suggestions.[[User:Vaishnavi Ananth\|Vaishnavi]]
		+	====Text Output====
		+	[[Engineering BioBrick vectors from BioBrick parts/Colony PCR protocol]]
		+	====Source Code====
		+	[[Engineering BioBrick vectors from BioBrick parts/Colony PCR protocol - source code]]

	[[Category:Protocol]]		[[Category:Protocol]]
	[[Category:Escherichia coli]]		[[Category:Escherichia coli]]
	[[Category:DNA]]		[[Category:DNA]]

Reshma P. Shetty at 16:00, 18 March 2008

2008-03-18T16:00:13Z

←Older revision		Revision as of 16:00, 18 March 2008
Line 7:		Line 7:
	*strip of PCR tubes		*strip of PCR tubes
	*[http://www.neb.com/nebecomm/products/productN3200.asp 2-log DNA ladder]		*[http://www.neb.com/nebecomm/products/productN3200.asp 2-log DNA ladder]
-	*0.8% E-Gel®	+	*0.8% E-Gel® from Invitrogen Corporation in Carlsbad, CA

	==Equipment==		==Equipment==

-	*DNA Engine OPTICON\texttrademark from MJ Research, Inc. (now Bio-Rad Laboratories, Inc., Hercules, CA).	+	*DNA Engine OPTICON\texttrademark from MJ Research, Inc. (now Bio-Rad Laboratories, Inc., Hercules, CA)
-	*E-Gel® PowerBase™ v.4 from Invitrogen Corporation in Carlsbad, CA.	+	*E-Gel® PowerBase™ v.4 from Invitrogen Corporation in Carlsbad, CA
	*Alpha Innotech FluoChem™ 880 gel imaging system		*Alpha Innotech FluoChem™ 880 gel imaging system

Reshma P. Shetty: /* Equipment */

2008-03-18T15:59:40Z

Equipment

←Older revision		Revision as of 15:59, 18 March 2008
Line 12:		Line 12:

	*DNA Engine OPTICON\texttrademark from MJ Research, Inc. (now Bio-Rad Laboratories, Inc., Hercules, CA).		*DNA Engine OPTICON\texttrademark from MJ Research, Inc. (now Bio-Rad Laboratories, Inc., Hercules, CA).
		+	*E-Gel® PowerBase™ v.4 from Invitrogen Corporation in Carlsbad, CA.
	*Alpha Innotech FluoChem™ 880 gel imaging system		*Alpha Innotech FluoChem™ 880 gel imaging system

Reshma P. Shetty at 15:57, 18 March 2008

2008-03-18T15:57:49Z

←Older revision		Revision as of 15:57, 18 March 2008
Line 6:		Line 6:
	*Deionized, sterile H<sub>2</sub>O		*Deionized, sterile H<sub>2</sub>O
	*strip of PCR tubes		*strip of PCR tubes
		+	*[http://www.neb.com/nebecomm/products/productN3200.asp 2-log DNA ladder]
		+	*0.8% E-Gel®

	==Equipment==		==Equipment==

	*DNA Engine OPTICON\texttrademark from MJ Research, Inc. (now Bio-Rad Laboratories, Inc., Hercules, CA).		*DNA Engine OPTICON\texttrademark from MJ Research, Inc. (now Bio-Rad Laboratories, Inc., Hercules, CA).
		+	*Alpha Innotech FluoChem™ 880 gel imaging system

	==Procedure==		==Procedure==
Line 30:		Line 33:
	#68°C for 20 mins		#68°C for 20 mins
	#4°C forever		#4°C forever
		+
		+	===Agarose gel electrophoresis===
		+
		+	#Dilute the reactions four-fold with water.
		+	#Perform an agarose gel electrophoresis of 20 μL of each diluted reaction using a 0.8% E-Gel®.
		+	#Also electrophorese 1 μg of 2-log DNA ladder to verify the length of each PCR product.
		+	#Image gel with 302 nm transilluminating ultraviolet light using an ethidium bromide camera filter and an exposure time of 614 milliseconds.

	[[Category:Protocol]]		[[Category:Protocol]]
	[[Category:Escherichia coli]]		[[Category:Escherichia coli]]
	[[Category:DNA]]		[[Category:DNA]]

Reshma P. Shetty: New page: ==Materials== [http://www.invitrogen.com/content/sfs/manuals/11306016.pdf PCR SuperMix High Fidelity] VF2 primer (`5'-TGCCACCTGACGTCTAAGAA-3'`) *VR primer (`5'-ATTACCGCC...`


2008-03-18T15:51:57Z
New page: ==Materials==  *[http://www.invitrogen.com/content/sfs/manuals/11306016.pdf PCR SuperMix High Fidelity] *VF2 primer (<code>5'-TGCCACCTGACGTCTAAGAA-3'</code>) *VR primer (<code>5'-ATTACCGCC...
New page
==Materials==



*[http://www.invitrogen.com/content/sfs/manuals/11306016.pdf PCR SuperMix High Fidelity]

*VF2 primer (<code>5'-TGCCACCTGACGTCTAAGAA-3'</code>)

*VR primer (<code>5'-ATTACCGCCTTTGAGTGAGC-3'</code>)

*Deionized, sterile H<sub>2</sub>O

*strip of PCR tubes



==Equipment==



*DNA Engine OPTICON\texttrademark from MJ Research, Inc. (now Bio-Rad Laboratories, Inc., Hercules, CA).



==Procedure==



===PCR mix===



*9 &mu;L PCR SuperMix High Fidelity

*6.25 pmoles VF2 primer

*6.25 pmoles VR primer

*1 &mu;L colony suspension 

**dilute 1 colony in 100 &mu;L water



===PCR conditions===



#95&deg;C for 15 minutes

#94&deg;C for 30 seconds

#62&deg;C for 30 seconds

#68&deg;C for 3.5 minutes

#Repeat 2-4 39 times.

#68&deg;C for 20 mins

#4&deg;C forever



[[Category:Protocol]]

[[Category:Escherichia coli]] 

[[Category:DNA]]

Engineering BioBrick vectors from BioBrick parts/Colony PCR - Revision history

Vaishnavi Ananth at 09:39, 19 November 2009

Vaishnavi Ananth at 06:23, 1 October 2009

Reshma P. Shetty at 16:00, 18 March 2008

Reshma P. Shetty: /* Equipment */

Reshma P. Shetty at 15:57, 18 March 2008

Reshma P. Shetty: New page: ==Materials== *[http://www.invitrogen.com/content/sfs/manuals/11306016.pdf PCR SuperMix High Fidelity] *VF2 primer (5'-TGCCACCTGACGTCTAAGAA-3') *VR primer (5'-ATTACCGCC...

Reshma P. Shetty: New page: ==Materials== [http://www.invitrogen.com/content/sfs/manuals/11306016.pdf PCR SuperMix High Fidelity] VF2 primer (`5'-TGCCACCTGACGTCTAAGAA-3'`) *VR primer (`5'-ATTACCGCC...`