Enzymes are the heart of each and every diagnostic test. Enzymes are the means by which we can track, study and defeat old and new infectious diseases. However, enzymes are globally distributed from centralized foundries and often require a cold chain to maintain function, restricting access. Poor access to reagents is exacerbated in times of supply chain disruptions such as those experienced during the COVID-19 pandemic.
The Open Bioeconomy Lab has developed the Molecular Diagnostics Toolkit to combat diagnostic enzyme access restrictions. The 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.
The Molecular Diagnostics Toolkit is available in two formats: E. coli glycerol stocks and purified nucleic acids. Please specify in the order notes if you need a particular format.
Some or all of these items are for use only as permitted by a research exemption.
Where can I find more information:
Special thanks to the Open Bioeconomy Lab for designing the Molecular Diagnostics Toolkit.
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 received the kit in the format of one plate, created after September 09, 2021, find the plate layout here.
If you have received the kit in the format of 2 different plates, check out the layout for Plate 1 and Plate 2. Briefly spin the plate at ≥500g to ensure that all DNA is at the bottom of the well before removing the foil seal.
The Molecular Diagnostics toolkit is available in a nucleic acid format. The parts in Plate 1 (or if only one plate, columns 1 and 2) are stored in pBuild, our standard cloning vector. The parts in Plate 2 (or if only one plate, columns 3 and 4) 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 | Freegenes ID |
|---|---|---|---|
| MERScontrol | MERS negative control | N/A | BBF10K_000004 |
| Pfu_Ssod7 HiFi | Pfu_Ssod7 HiFi - High Fidelity DNA polymerase | BBF10K_000006 | |
| gp32 | gp32 | BBF10K_000013 | |
| Ecoli_opt_RNAseInhib | E coli optimized RNAse Inhibitor | BBF10K_000014 | |
| HIV-RT | Human Immunodeficiency Virus Reverse Transcriptase | BBF10K_003500 | |
| OMMLV | Moloney Murine Leukemia Virus Reverse Transcriptase | BBF10K_003501 | |
| OpenVent | OpenVent DNA polymerase | BBF10K_003502 | |
| RNAseInhib | RNAse Inhibitor | BBF10K_003503 | |
| EcPPA Pyrophosphatase | Inorganic pyrophosphatase from E.coli | BBF10K_003504 | |
| T7_Orf1A_1 | SARS-CoV-2 Orf1A positive control | N/A | BBF10K_003505 |
| T7_Orf1A_2 | SARS-CoV-2 Orf1A 2 positive control | N/A | BBF10K_003506 |
| T7_Orf1B_3 | SARS-CoV-2 Orf1B positive control | N/A | BBF10K_003507 |
| T7_Orf1E_4 | SARS-CoV-2 Orf1E positive control | N/A | BBF10K_003508 |
| T7_OrfN_5 | SARS-CoV-2 OrfN positive control | N/A | BBF10K_003509 |
| Sars_NegativeControl | SARS-CoV-2 negative control | N/A | BBF10K_003510 |
| Bst-LF | Bst-LF | BBF10K_003564 | |
| Bsu | Bsu | BBF10K_003565 | |
| UvsX | UvsX | BBF10K_003566 | |
| PBCV-1_ligase | PBCV-1_ligase | BBF10K_003567 | |
| TthRNAseH | TthRNAseH | BBF10K_003568 | |
| EcRNAseH | EcRNAseH | BBF10K_003569 | |
| TEV | TEV | BBF10K_003570 |
Download all of this information as a CSV from our GitHub.
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 | Country |
|---|---|---|
| Kefentse Arnold Tumedi | tumediak {at} yahoo {dot} com | Botswana |
| Ahmed Atef | ahmed_atefaig2 {at} yahoo {dot} com | United States |
| Dong Nguyen Tam | NA | Vietnam |
| Johan Sosa | johan {at} thefunquantum {dot} com | United States |
| Vitaliy Strochkov | NA | Kazakhstan |
| Deepak Saini | deepaksaini {at} iisc {dot} ac {dot} in | India |
| Rodolfo Marcolino | rodolfo.marcolino {at} me {dot} com | Argentina |
| Ferdinand Molnár | NA | Kazakhstan |
| Zhiyong Lei | zlei {at} umcutrecht {dot} nl | Netherlands |
| Xingyue jiang | karma5781 {at} 21cn {dot} com | China |
| Esteban Erben | eerben {at} iib {dot} unsam {dot} edu {dot} ar | Argentina |
| Louise Dalgaard | ltd {at} ruc {dot} dk | Denmark |
| Ingrid Lorena Jaramillo | Lojadel2 {at} gmail {dot} com | Colombia |
| Ahmed Hegazy | a.hegazy {at} biotec {dot} rwth-aachen {dot} de | Germany |
| Jose David Rosales | jdrr55 {at} yahoo {dot} com https://www {dot} linkedin {dot} com/in/david-rosales-574a7151/ | Venezuela |
| Jennifer Hsieh | j.hsieh {at} kurabiotech {dot} com | Chile |
| Leandro Oliveira | leandro.licursi {at} ufv {dot} br | United Kingdom |
| TING-WEI LIN | weitinglin66 {at} gmail {dot} com | Taiwan |
| Matthew Phanchana | matthew.pha {at} mahidol {dot} edu | Thailand |
| Ahmed Atef | ahmed_atefaig2 {at} yahoo {dot} com | Saudi Arabia |
| Simran Kushwaha | https://www.bits-pilani.ac.in/pilani/biologicalScience/Faculty | India |
| Emin Bursa | NA | United States |
| Jimmy Gledson Hayden Linhares | haydenlinhars {at} gmail {dot} com | Brazil |
| Keoni Gandall | koeng101 {at} gmail {dot} com | United States |
| James Altamirano | James_altam {at} yahoo {dot} com | United States |
| Jordan Gonzalez | jgonzalez {at} thecitizensciencelab {dot} org | United States |
| Jan Studeny | jan.studeny {at} aalto {dot} fi | Finland |
| Carlos Ueira-Vieira | ueira {at} ufu {dot} br | Brazil |
| Carlos Alexandre Breyer | carlosbreyer {at} gmail {dot} com | Brazil |
| Joey Riepsaame | joey.riepsaame {at} path {dot} ox {dot} ac {dot} uk | United Kingdom |
| Wei-Min Chang | WeiMinChang {at} tmu {dot} edu {dot} tw | Taiwan |
| Amjed Alsultan | amjed.talib {at} qu {dot} edu {dot} iq | Iraq |
| Majed Alghamdi | mafa2 {at} le {dot} ac {dot} uk | United Kingdom |
| raghav sridhar | raghav {at} cambrianbioworks {dot} com | India |
| Han Teng Wong | wong_han_teng {at} imcb {dot} a-star {dot} edu {dot} sg | Singapore |
| Kenneth Frimpong | ken.frimpong {at} thrivusinstitute {dot} edu {dot} gh | Ghana |
| Pratik Vyas | iampratikvyas {at} gamil {dot} com | India |
| Amitabha Majumdar | Mamitava {at} nccs {dot} res {dot} in | India |
| Xavier Coadic | xcoadic {at} protonmail {dot} com | France |
Download all of this information as a CSV from our GitHub.