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 shipped as purified and dried down plasmid DNA stained with Cresol Red in one 96-well plate. Each well contains approximately 50 ng of DNA. Cresol Red will not impact plasmid transformation.
Some or all of these items are for use only as permitted by a research exemption
Current plate map can be found here.
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 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 |
|---|---|---|
| Jordan Gonzalez | jgonzalez {at} thecitizensciencelab {dot} org | United States |
| Jose David Rosales | jdrr55 {at} yahoo {dot} com https://www {dot} linkedin {dot} com/in/david-rosales-574a7151/ | Venezuela |
| Louise Dalgaard | ltd {at} ruc {dot} dk | Denmark |
| Jan Studeny | jan.studeny {at} aalto {dot} fi | Finland |
| Tri Ngo | NA | United States |
| Wei-Min Chang | WeiMinChang {at} tmu {dot} edu {dot} tw | Taiwan |
| Amjed Alsultan | amjed.talib {at} qu {dot} edu {dot} iq | Iraq |
| Deepak Saini | deepaksaini {at} iisc {dot} ac {dot} in | India |
| Ben Davis | bed58 {at} aber {dot} ac {dot} uk | United Kingdom |
| Norhan Hassan | norhan.hassan {at} kaust {dot} edu {dot} sa | Saudi Arabia |
| Rodolfo Marcolino | rodolfo.marcolino {at} me {dot} com | Argentina |
| Vitaliy Strochkov | NA | Kazakhstan |
| Tomasz Jurkowski | jurkowskit {at} cardiff {dot} ac {dot} uk | United Kingdom |
| Dong Nguyen Tam | NA | Vietnam |
| Ferdinand Molnár | NA | Kazakhstan |
| Cihan Aydin | cihan.aydin {at} medeniyet {dot} edu {dot} tr | Turkey |
| Zhiyong Lei | zlei {at} umcutrecht {dot} nl | Netherlands |
| Christopher Kinyanjui Kariuki | christopherkinyanjui.kariuki {at} kuleuven {dot} be | Belgium |
| Jimmy Gledson Hayden Linhares | haydenlinhars {at} gmail {dot} com | Brazil |
| Vincent Paris | NA | United States |
| Ahmed Atef | ahmed_atefaig2 {at} yahoo {dot} com | Saudi Arabia |
| Stephen Klusza | smklusza {at} gmail {dot} com | United States |
| Ahmad Suparmin | ahmad.suparmin {at} ugm {dot} ac {dot} id | Indonesia |
| Simran Kushwaha | https://www.bits-pilani.ac.in/pilani/biologicalScience/Faculty | India |
| Jennifer Hsieh | j.hsieh {at} kurabiotech {dot} com | Chile |
| Ganesh Prasad | ganeshmayya {at} gmail {dot} com | India |
| Kenneth Frimpong | ken.frimpong {at} thrivusinstitute {dot} edu {dot} gh | Ghana |
| Johan Sosa | johan {at} thefunquantum {dot} com | United States |
| Amitabha Majumdar | Mamitava {at} nccs {dot} res {dot} in | India |
| James Altamirano | James_altam {at} yahoo {dot} com | United States |
| Jose David Rosales | https://www.linkedin.com/in/david-rosales-574a7151/ | United States |
| Ahmed Hegazy | a.hegazy {at} biotec {dot} rwth-aachen {dot} de | Germany |
| Ahmed Atef | ahmed_atefaig2 {at} yahoo {dot} com | United States |
| Long Nguyen | Lnguyen2 {at} ufl {dot} edu | United States |
| Bingbing Wan | wanb {at} sjtu {dot} edu {dot} cn | China |
| Andrei Berasneu | andreyberesnevson {at} gmail {dot} com | Poland |
| Pratik Vyas | iampratikvyas {at} gamil {dot} com | India |
| Ingrid Lorena Jaramillo | Lojadel2 {at} gmail {dot} com | Colombia |
| Emin Bursa | NA | United States |
| Xingyue jiang | karma5781 {at} 21cn {dot} com | China |
| TING-WEI LIN | weitinglin66 {at} gmail {dot} com | Taiwan |
| Stephen Klusza | StephenKlusza {at} clayton {dot} edu | United States |
| Irfan Hussain | irfan {at} bs {dot} qau {dot} edu {dot} pk | Pakistan |
| Shibichakravarthy Kannan | shibi.kannan {at} gmail {dot} com | India |
| Carlos Alexandre Breyer | carlosbreyer {at} gmail {dot} com | Brazil |
| Codruta Ignea | codruta.ignea {at} mcgill {dot} ca | Canada |
| Keoni Gandall | koeng101 {at} gmail {dot} com | United States |
| Xavier Coadic | xcoadic {at} protonmail {dot} com | France |
| Joey Riepsaame | joey.riepsaame {at} path {dot} ox {dot} ac {dot} uk | United Kingdom |
| Matthew Phanchana | matthew.pha {at} mahidol {dot} edu | Thailand |
| Maira Goytia | mgoytia {at} spelman {dot} edu | United States |
| Benjamin Loang | benjamin.liang {at} uq {dot} edu {dot} au | Australia |
| Joao Vitor Dutra Molino | candidomolino {at} gmail {dot} com | United States |
| Esteban Erben | eerben {at} iib {dot} unsam {dot} edu {dot} ar | Argentina |
| Han Teng Wong | wong_han_teng {at} imcb {dot} a-star {dot} edu {dot} sg | Singapore |
| Majed Alghamdi | mafa2 {at} le {dot} ac {dot} uk | United Kingdom |
| Nadia Odaliz Chamana Chura | nadia.chamana {at} utec {dot} edu {dot} pe | France |
| Leandro Oliveira | leandro.licursi {at} ufv {dot} br | United Kingdom |
| Carlos Ueira-Vieira | ueira {at} ufu {dot} br | Brazil |
| raghav sridhar | raghav {at} cambrianbioworks {dot} com | India |
| Marko Komlosh | markokomlosh {at} gmail {dot} com | Serbia |
| Kefentse Arnold Tumedi | tumediak {at} yahoo {dot} com | Botswana |
| Krishna Gupta | gupta-krishna {at} ipfdd {dot} de | Germany |
Download all of this information as a CSV from our GitHub.