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| <center>'''Amplified Insert Assembly'''</center> | | <center>'''Amplified Insert Assembly'''</center> |
| ==Overview== | | ==Overview== |
| | | This is a lab specific protocol. For more information on the benefits of Amplified Insert Assembly see the [[Amplified Insert Assembly| consensus protocol. |
| Amplified Insert Assembly is a method of "BioBricking" two biological parts (i.e. pieces of DNA) together and was developed by [[User:Michael A. Speer|Mike Speer]] and Dr. Tom Richard. 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. In comparison to [[Synthetic Biology:BioBricks/3A assembly|3A assembly]], this method can take up to two hours longer; however, the added time spent at the bench is minimal. Major benefits of this assembly method over other bio-brick assembly methods include:
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| *no need for [[Richard Lab:Agarose Gel Electrophoresis|gel electrophoresis]] or [[DNA Gel extraction|gel extraction]].
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| *the ability to insert small (i.e. invisible on a gel) parts.
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| *no need to use [[Synthetic Biology:BioBricks/3A assembly|multiple antibiotic resistances]].
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| *no having to make construction vectors.
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| *really low background (99% of colonies are correct)
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| **This means less sequencing
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| *Easy transformation (use homemade competent cells)
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| *Less culturing
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| **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.
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| 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: |
