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From 2010.igem.org

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	{{:Team:uOttawa/Templates/header}}		{{:Team:uOttawa/Templates/header}}
-		+	=Protocols=
-	{|align="justify"	+	==Dip and swirl cloning==
-	|You can write a background of your team here.  Give us a background of your team, the members, etc.  Or tell us more about something of your choosing.	+	This protocol is essentially the same as the Standard BioBrick Cloning protocol used in our lab with the exception of using a minimal amount of restriction enzymes.
-	|[[Image:uOttawa_logo.png|200px|right|frame]]	+	*'''Restriction Digest'''
-	|-	+	##Add 0.3 μl of 10X NEBbuffer 2
-	|	+	##Add 0.3 μl 0f 10X BSA
-	''Tell us more about your project.  Give us background.  Use this is the abstract of your project.  Be descriptive but concise (1-2 paragraphs)''	+	##Add 30 ng of DNA (plasmid or PCR product)
-	|[[Image:uOttawa_team.png|right|frame|Your team picture]]	+	##Dip the pipette tip in the enzyme solution and move it around (swirl). Place the tip in your reaction mixture, move it around (swirl) again, and rinse it.
-	|-	+	##Add water to a final volume of 3 μl.
-	|	+	*The remaining steps of the protocol are identical to that of the Standard BioBrick Cloning protocol.
-	|align="center"|[[Team:uOttawa | Team Example]]	+	'''N.B.''' This protocol is particularly suitable for when the enzyme solution has almost run out, but you still need to perform a number of digestions. This protocol is longer than the Standard BioBrick Cloning protocol. In addition, if the level of the enzyme solution is high, the capillarity action causes more than enough of the enzyme to enter the tip, hence, rendering this approach pointless.
-	|}	+
-		+
-	<!--- The Mission, Experiments --->	+
-		+
-	{| style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"	+
-	!align="center"|[[Team:uOttawa|Home]]	+
-	!align="center"|[[Team:uOttawa/Team|Team]]	+
-	!align="center"|[https://igem.org/Team.cgi?year=2010&team_name=uOttawa Official Team Profile]	+
-	!align="center"|[[Team:uOttawa/Project|Project]]	+
-	!align="center"|[[Team:uOttawa/Parts|Parts Submitted to the Registry]]	+
-	!align="center"|[[Team:uOttawa/Modeling|Modeling]]	+
-	!align="center"|[[Team:uOttawa/Notebook|Notebook]]	+
-	!align="center"|[[Team:uOttawa/Safety|Safety]]	+
-	|}	+
-		+
-		+
-	==Notebook==	+
-		+
-	You should make use of the calendar feature on the wiki and start a lab notebook. This may be looked at by the judges to see how your work progressed throughout the summer. It is a very useful organizational tool as well.	+

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Revision as of 06:11, 27 October 2010

Protocols

Dip and swirl cloning

This protocol is essentially the same as the Standard BioBrick Cloning protocol used in our lab with the exception of using a minimal amount of restriction enzymes.

• Restriction Digest

1. 1. Add 0.3 μl of 10X NEBbuffer 2
   2. Add 0.3 μl 0f 10X BSA
   3. Add 30 ng of DNA (plasmid or PCR product)
   4. Dip the pipette tip in the enzyme solution and move it around (swirl). Place the tip in your reaction mixture, move it around (swirl) again, and rinse it.
   5. Add water to a final volume of 3 μl.

• The remaining steps of the protocol are identical to that of the Standard BioBrick Cloning protocol.

N.B. This protocol is particularly suitable for when the enzyme solution has almost run out, but you still need to perform a number of digestions. This protocol is longer than the Standard BioBrick Cloning protocol. In addition, if the level of the enzyme solution is high, the capillarity action causes more than enough of the enzyme to enter the tip, hence, rendering this approach pointless.