Experiments
Project Management
Experiments: Overview
- Transcriptome analysis of T. solium
- RNA Isolation
- RNA sequencing
- Bioinformatics
- Switch Synthesis
- Extension PCR
- Nested PCR
- Colony PCR
- Gel Electrophoresis
- DNA Clean-Up
- Toehold Switch Sensor Test – Testing Pipeline
- High troughput screening in liquid medium
- Reliable cell free expression on cellulose membranes
- Optimizing the Screening Pipeline
- Other Experiments
- Preparation of nucleic acids (DNA preparation, Restriction digest/ DNA ligation, in vitro transcription, RNA purification)
- Preparation of sensors for iGEM Submission (Mutagenesis & Cloning)
Please find detailed protocols under the "Protocol" Section
Transcriptome Analysis of T. solium
RNA Isolation
Toehold switch sensors are based on synthetic biology. Essentially, they are RNA molecules which code for a reporter protein. They consist of a specific toehold sequence, a ribosome-binding site (which is important for the production of proteins) and a sequence for the reporter protein. The reporter protein can only be produced if the sensor has bonded with its specific target RNA sequence [4]. When producing the sensors, we can select both the toehold sequence and the reporter protein with complete flexibility to match our needs. In this way we can fashion our Wormspotter so that it only sends a desired signal when it binds to RNA molecules specific to T. solium. We are planning to use T. solium-specific RNA sequences for the toehold sequence and beta-Galactosidase as a reporter protein.
RNA Sequencing
Toehold switch sensors are based on synthetic biology. Essentially, they are RNA molecules which code for a reporter protein. They consist of a specific toehold sequence, a ribosome-binding site (which is important for the production of proteins) and a sequence for the reporter protein. The reporter protein can only be produced if the sensor has bonded with its specific target RNA sequence [4]. When producing the sensors, we can select both the toehold sequence and the reporter protein with complete flexibility to match our needs. In this way we can fashion our Wormspotter so that it only sends a desired signal when it binds to RNA molecules specific to T. solium. We are planning to use T. solium-specific RNA sequences for the toehold sequence and beta-Galactosidase as a reporter protein.