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User:Meagan/Assembly standards
From 2008.igem.org
(New page: People in community have developed various cloning techniques/assembly standards. iGEM supports three so far. (But there are a lot more being worked on) Why would you want to use stand...) |
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* more reliable | * more reliable | ||
* less time consuming | * less time consuming | ||
| - | * separate function from assembly | + | * each resulting part fits the same assembly scheme as was used to create it |
| + | * can separate function from assembly | ||
* robotic assembly | * robotic assembly | ||
| - | === BioBrick™ Assembly <small>(TKnight)</small> === | + | === <font size=4>BioBrick™ Assembly </font><small>(TKnight)</small> === |
| - | BioBrick™ assembly is the original assembly standard created by Tom Knight. It was first described in his paper on [http://hdl.handle.net/1721.1/21168 Idempotent Vector Design for Standard Assembly of Biobricks]. Since publication of that paper, this assembly standard has been adopted by Tom Knights lab the Registry of standard Biological Parts, iGEM, and many labs in the synthetic biology (based on standard biological parts) community. | + | BioBrick™ assembly is the original assembly standard created by Tom Knight. It was first described in his paper on [http://hdl.handle.net/1721.1/21168 Idempotent Vector Design for Standard Assembly of Biobricks™]. Since publication of that paper, this assembly standard has been adopted by Tom Knights lab the Registry of standard Biological Parts, iGEM, and many labs in the synthetic biology (based on standard biological parts) community. |
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| + | Various methods: | ||
| + | * standard assembly | ||
| + | * 3-antibiotic assembly | ||
| - | |||
| + | === <font size=4>FusionBricks </font><small>(Silver Lab)</small> === | ||
| + | Developed by Pam Silver, Ira Phillips, and the rest of the Silver lab at Harvard. They found that the original BioBrick™ scheme was not compatible when considering proteins. The scar that is created by the BioBrick™ assembly method creates a frame shift when translating to a protein. | ||
| + | The FusionBricks method allows the frame to be maintained across the scar by adding a different number of spacer nucleotides on either side of the part sequence. | ||
| + | IMAGE OF SCHEME | ||
| - | === BioBricksExtreme Assembly <small>(JCAnderson)</small> === | + | |
| + | Other features/requirements of using FusionBricks: | ||
| + | * XbaI and SpeI restriction sites are directly next to the part | ||
| + | * Parts must not begin with a T or a C nucleotide | ||
| + | * BioBrick™ parts can be combined with FusionBrick parts | ||
| + | |||
| + | |||
| + | |||
| + | Supported plasmids: | ||
| + | * same as the BioBrick™ supported plasmids (see above) | ||
| + | |||
| + | |||
| + | |||
| + | === <font size=4>BioBricksExtreme Assembly </font><small>(JCAnderson)</small> === | ||
Revision as of 18:59, 23 April 2008
People in community have developed various cloning techniques/assembly standards. iGEM supports three so far. (But there are a lot more being worked on)
Why would you want to use standard biological parts and an assembly standard?
- easier than traditional cloning techniques
- more reliable
- less time consuming
- each resulting part fits the same assembly scheme as was used to create it
- can separate function from assembly
- robotic assembly
BioBrick™ Assembly (TKnight)
BioBrick™ assembly is the original assembly standard created by Tom Knight. It was first described in his paper on Idempotent Vector Design for Standard Assembly of Biobricks™. Since publication of that paper, this assembly standard has been adopted by Tom Knights lab the Registry of standard Biological Parts, iGEM, and many labs in the synthetic biology (based on standard biological parts) community.
Supported plasmids:
- pSBxxx series, e.g.:
- pSB1AC3
Restriction enzyme scheme:
- EcoRI (Prefix)
- XbaI (Prefix)
- SpeI (Suffix)
- PstI (Suffix)
Various methods:
- standard assembly
- 3-antibiotic assembly
FusionBricks (Silver Lab)
Developed by Pam Silver, Ira Phillips, and the rest of the Silver lab at Harvard. They found that the original BioBrick™ scheme was not compatible when considering proteins. The scar that is created by the BioBrick™ assembly method creates a frame shift when translating to a protein.
The FusionBricks method allows the frame to be maintained across the scar by adding a different number of spacer nucleotides on either side of the part sequence.
IMAGE OF SCHEME
Other features/requirements of using FusionBricks:
- XbaI and SpeI restriction sites are directly next to the part
- Parts must not begin with a T or a C nucleotide
- BioBrick™ parts can be combined with FusionBrick parts
Supported plasmids:
- same as the BioBrick™ supported plasmids (see above)
