Difference between revisions of "Part:BBa K1614016:Design"
(→Design Notes) |
|||
| Line 7: | Line 7: | ||
===Design Notes=== | ===Design Notes=== | ||
| − | Cloned without restriction enzymes to avoid issues occurring from conserved cut sites. For more details see BBF RFC 110. | + | Cloned without restriction enzymes to avoid issues occurring from conserved cut sites. For more details see BBF RFC 110. |
| − | + | The part was designed to live track the <i>in vitro</i> transcription of any RNA of interest (ROI). Every ROI can be cloned into this part behind the T7 promotor and in front of the hammerhead ribozyme (HHR) using cloning techniques proposed by our team in BBF RFC 110. | |
| − | + | This way it is possible so track in real-time how much ROI is transcribed. On top of that it increases the amount of correctly transcribed ROI and results in a defined 3' end as the HHR cleaves off at the defined position. This makes universal measurement device. | |
===Source=== | ===Source=== | ||
Latest revision as of 13:51, 20 September 2015
Hammerhead Ribozyme and Malchite Green Aptamer in BBF RFC 110 transcription cassette
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 85
Illegal BamHI site found at 117 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 124
Illegal NgoMIV site found at 153 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
Cloned without restriction enzymes to avoid issues occurring from conserved cut sites. For more details see BBF RFC 110. The part was designed to live track the in vitro transcription of any RNA of interest (ROI). Every ROI can be cloned into this part behind the T7 promotor and in front of the hammerhead ribozyme (HHR) using cloning techniques proposed by our team in BBF RFC 110. This way it is possible so track in real-time how much ROI is transcribed. On top of that it increases the amount of correctly transcribed ROI and results in a defined 3' end as the HHR cleaves off at the defined position. This makes universal measurement device.
Source
Synthetic, Malchit Green aptamer (Grate, D., and Wilson, C. (1999). Laser-mediated, site-specific inactivation of RNA transcripts. Proceedings of the National Academy of Sciences of the United States of America 96, 6131-6136.) HHR (Prody, G.A., Bakos, J.T., Buzayan, J.M., Schneider, I.R., and Bruening, G. (1986). Autolytic processing of dimeric plant virus satellite RNA. Science 231, 1577-1580)