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Richard Lab:Amplified insert assembly: Difference between revisions
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Richard Lab:Amplified insert assembly (view source)
Revision as of 15:18, 25 January 2011
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==Overview== | ==Overview== | ||
This is what [[User:Michael A. Speer|Mike]] calls "New Standard Assembly" and is a method of "bio-bricking" two biological parts (i.e. pieces of DNA) together. For more information on bio-bricking see [http://partsregistry.org/Help:Contents this link]. This method combines the ease and speed of 3A assembly with the | This is what [[User:Michael A. Speer|Mike]] calls "New Standard Assembly" and is a method of "bio-bricking" two biological parts (i.e. pieces of DNA) together. For more information on bio-bricking see [http://partsregistry.org/Help:Contents this link]. This method combines the ease and speed of 3A assembly with the reliability of standard assembly. Major benefits of this assembly method over other assembly methods include: | ||
*no need for [[Richard Lab:Agarose Gel Electrophoresis|gel electrophoresis]] or [[DNA Gel extraction|gel extraction]]. | *no need for [[Richard Lab:Agarose Gel Electrophoresis|gel electrophoresis]] or [[DNA Gel extraction|gel extraction]]. | ||
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The two parts you want to assemble will be labeled "insert" and "vector" and will be initially contained on separate plasmids. The eventual goal of assembly is to get these parts on the same plasmid next to one another. | The two parts you want to assemble will be labeled "insert" and "vector" and will be initially contained on separate plasmids. The eventual goal of assembly is to get these parts on the same plasmid next to one another. | ||
==Materials== | |||
===General=== | |||
*Pipettors | |||
*Microcentrifuge | |||
*Water baths | |||
*Thermocycler | |||
*Electroporator | |||
*Selective media plates | |||
===Enzymes=== | |||
*EcoRI Restriction Endonuclease | |||
*XbaI Restriction Endonuclease | |||
*SpeI Restriction Endonuclease | |||
*PstI Restriction Endonuclease | |||
*DpnI Restriction Endonuclease | |||
*Vent DNA Polymerase | |||
*Antarctic Phosphatase | |||
*T4 Ligase | |||
==Procedure== | ==Procedure== | ||
