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Richard Lab:Amplified insert assembly: Difference between revisions
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Richard Lab:Amplified insert assembly (view source)
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==Overview== | ==Overview== | ||
This is what [[User:Michael A. Speer|Mike]] calls "New Standard Assembly" and is a method of " | This is what [[User:Michael A. Speer|Mike]] calls "New Standard Assembly" and is a method of "BioBricking" 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 bio-brick 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]]. | ||
*the ability to insert small (i.e. invisible on a gel) parts. | *the ability to insert small (i.e. invisible on a gel) parts. | ||
*no need to | *no need to use [[Synthetic Biology:BioBricks/3A assembly|multiple antibiotic resistances]]. | ||
*no having to make construction vectors. | *no having to make construction vectors. | ||
*really low background transformation | *really low background (99% of colonies are correct) | ||
**This means less sequencing | |||
*Easy transformation (use homemade competent cells) | |||
*Less culturing | |||
**This is because one plasmid prep can supply many PCR inserts. So common parts (i.e. promoters and RBSs) can be used over and over again. | |||
This protocol is typically used to do bio-brick assembly with restriction sites consisting of the following configuration: | This protocol is typically used to do bio-brick assembly with restriction sites consisting of the following configuration: | ||
