Difference between revisions of "Part:BBa K1614002:Experience"
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===Applications of BBa_K1614002=== | ===Applications of BBa_K1614002=== | ||
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| − | + | A highly relevant series of parts that is based upon BBa_K1614002 are all parts associated to our RFC (as also described above). In our RFC 110, we describe a standardized procedure for the production of small functional RNA. The storage of the constructs, as well as the integration into the <i>in vitro</i> transcription - this cassette is suggested to be performed in BBa_K1614002. Therefore, this part is also central to our work and by including it into the registry, we want to enable the entire community of synthetic biology to generate small functional RNA in a standardized, yet versatile way. | |
| − | A highly relevant series of parts that is based upon BBa_K1614002 are all parts associated to our RFC (as also described above). In our RFC 110, we describe a standardized procedure for the production of small functional RNA. The storage of the constructs, as well as the integration into the <i>in vitro</i> transcription - cassette is suggested to be performed in BBa_K1614002. Therefore, this part is also central to our work and by including it into the registry, we want to enable the entire community of synthetic biology to generate small functional RNA in a standardized, yet versatile way. | + | |
</p> | </p> | ||
[[File:HeidelbergPWiGEM2015_RFC.png|thumb|General mechanism of the RFC which is based on BioBrick BBa_K1614002]] | [[File:HeidelbergPWiGEM2015_RFC.png|thumb|General mechanism of the RFC which is based on BioBrick BBa_K1614002]] | ||
<p> | <p> | ||
| − | For the work with the BBF RFC proposed here, we suggest the workflow described in Figure 1. A more detailed description of the required protocols can be found in the file of our RFC. | + | For the work with the BBF RFC 110 proposed here, we suggest the workflow described in Figure 1. A more detailed description of the required protocols can be found in the file of our RFC. |
</p> | </p> | ||
<p> | <p> | ||
| − | After linearization of BBa_K1614002 with standard primers (BBa_1614003 and BBa_1614004), a synthesized insert with homologous overhangs is cloned in via CPE Cloning or Gibson Assembly. As BBa_K1614002 already contains the T7 promoter and the HDV-ribozyme for defined ends of the transcript, the sequence of the desired RNA can be inserted in a standardized way and the extraction of the DNA-template of the functional RNA can be extracted with standard primers (BBa_K1614005 and BBa_1614006) and transcribed in vitro. In this process, quality of the in vitro transcription can be easily monitored and conditions can be optimized. | + | After linearization of BBa_K1614002 with standard primers (BBa_1614003 and BBa_1614004), a synthesized insert with homologous overhangs is cloned in via CPE Cloning or Gibson Assembly. As BBa_K1614002 already contains the T7 promoter and the HDV-ribozyme for defined ends of the transcript, the sequence of the desired RNA can be inserted in a standardized way and the extraction of the DNA-template of the functional RNA can be extracted with standard primers (BBa_K1614005 and BBa_1614006) and transcribed ''in vitro''. In this process, quality of the ''in vitro'' transcription can be easily monitored and conditions can be optimized. |
</p> | </p> | ||
<p> | <p> | ||
Latest revision as of 18:52, 20 September 2015
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how you used this part and how it worked out.
Applications of BBa_K1614002
A highly relevant series of parts that is based upon BBa_K1614002 are all parts associated to our RFC (as also described above). In our RFC 110, we describe a standardized procedure for the production of small functional RNA. The storage of the constructs, as well as the integration into the in vitro transcription - this cassette is suggested to be performed in BBa_K1614002. Therefore, this part is also central to our work and by including it into the registry, we want to enable the entire community of synthetic biology to generate small functional RNA in a standardized, yet versatile way.
For the work with the BBF RFC 110 proposed here, we suggest the workflow described in Figure 1. A more detailed description of the required protocols can be found in the file of our RFC.
After linearization of BBa_K1614002 with standard primers (BBa_1614003 and BBa_1614004), a synthesized insert with homologous overhangs is cloned in via CPE Cloning or Gibson Assembly. As BBa_K1614002 already contains the T7 promoter and the HDV-ribozyme for defined ends of the transcript, the sequence of the desired RNA can be inserted in a standardized way and the extraction of the DNA-template of the functional RNA can be extracted with standard primers (BBa_K1614005 and BBa_1614006) and transcribed in vitro. In this process, quality of the in vitro transcription can be easily monitored and conditions can be optimized.
After self-splicing of the HDV-ribozyme has occurred, the functional RNA can be extracted from the reaction mixture and purified via Polyacrylamide Gel Electrophoresis (PAGE).
In order to create novel aptamers or aptamer-switchable ribozymes, we here reference to the possibility to create new aptamers via the software MAWS (Making Aptamers Without SELEX) http://2015.igem.org/Team:Heidelberg/software/maws and the software JAWS (Joining Aptamers Without SELEX) http://2015.igem.org/Team:Heidelberg/software/jaws.
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