Enzymes are the heart of each and every diagnostic test. Enzymes are the means we can track, study and defeat old and new infectious diseases. However, enzymes are globally distributed from centralized foundries and, most often, require a cold chain for stability restricting their access to the many, even more in times of supply chain disruptions like is happening in the current COVID-19 pandemic.
The Molecular Diagnostics Toolkit includes:
- IPTG inducible, T7 promoter driven ready-to-use expression cassettes to produce essential enzymes used in diagnostic among which RT, Bst-LF, RPA enzymes, RNA inhibitors. Enzymes are His-tagged and cassettes design are based on literature and experimental evidence
- A set of constructs for the production of Sars-CoV-2 RNA positive and negative controls
What can it be used for:
The collection wants to provide researchers worldwide with an open source toolkit to produce their own enzymes, streamlining their research in diagnostics irrespective of supply chain disruptions.
The expression cassettes in this collection have been shown to work in literature. For distribution purposes we had to clone the newly synthesized cassettes in a novel off-patent vector backbone. The expression from this new vector has not been validated yet.
This product is made available under the unilateral OpenMTA.
All genes are present on the plate in E. coli glycerol stocks. Some or all of these items are for use only as permitted by a research exemption.
Where can I find more information:
If you would like to design your own expression cassette, changing promoter, tag, reporter gene or terminator have a look at our Expression Toolkit collection.
If you have already received the kit, check out the layout for Plate 1 and Plate 2.
The parts in Plate 1 are stored in pBuild, our standard cloning vector. The parts in Plate 2 are stored in pInducible, an inducible-copy backbone with a kanamycin selection marker. To induce pInducible’s high copy origin of replication, add IPTG to the mid-log-phase culture to a final concentration of 1mM. Allow cells to grow to saturation, then miniprep DNA.
Genes
| Gene | Name | NCBI ID |
|---|---|---|
| MERScontrol | MERS negative control | N/A |
| Pfu_Ssod7 HiFi | Pfu_Ssod7 HiFi - High Fidelity DNA polymerase | |
| gp32 | gp32 | |
| Ecoli_opt_RNAseInhib | E coli optimized RNAse Inhibitor | |
| HIV-RT | Human Immunodeficiency Virus Reverse Transcriptase | |
| OMMLV | Moloney Murine Leukemia Virus Reverse Transcriptase | |
| OpenVent | OpenVent DNA polymerase | |
| RNAseInhib | RNAse Inhibitor | |
| EcPPA Pyrophosphatase | Inorganic pyrophosphatase from E.coli | |
| T7_Orf1A_1 | SARS-CoV-2 Orf1A positive control | N/A |
| T7_Orf1A_2 | SARS-CoV-2 Orf1A 2 positive control | N/A |
| T7_Orf1B_3 | SARS-CoV-2 Orf1B positive control | N/A |
| T7_Orf1E_4 | SARS-CoV-2 Orf1E positive control | N/A |
| T7_OrfN_5 | SARS-CoV-2 OrfN positive control | N/A |
| Sars_NegativeControl | SARS-CoV-2 negative control | N/A |
| Bst-LF | Bst-LF | |
| Bsu | Bsu | |
| UvsX | UvsX | |
| PBCV-1_ligase | PBCV-1_ligase | |
| TthRNAseH | TthRNAseH | |
| EcRNAseH | EcRNAseH | |
| TEV | TEV |
Bionet
The bionet enables open peer-peer exchange of functional biomaterials and associated data. This product may also be available from bionet nodes that are more convenient to you. Here are other bionet nodes who may be willing to provide you this specific product.| Name | Contact |
|---|---|
| Louise Dalgaard | ltd {at} ruc {dot} dk |
| Vitaliy Strochkov | NA |
| Amitabha Majumdar | Mamitava {at} nccs {dot} res {dot} in |
| Dong Nguyen Tam | NA |
| Amjed Alsultan | amjed.talib {at} qu {dot} edu {dot} iq |
| Matthew Phanchana | matthew.pha {at} mahidol {dot} edu |
| Jordan Gonzalez | jgonzalez {at} thecitizensciencelab {dot} org |
| Keoni Gandall | koeng101 {at} gmail {dot} com |