Molecular Diagnostics Toolkit

Molecular Diagnostics Toolkit

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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. Cresol Red will not impact plasmid transformation.

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, 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
Xingyue jiang karma5781 {at} 21cn {dot} com China
Christopher Kinyanjui Kariuki christopherkinyanjui.kariuki {at} kuleuven {dot} be Belgium
Carlos Alexandre Breyer carlosbreyer {at} gmail {dot} com Brazil
Ferdinand Molnár NA Kazakhstan
James Altamirano James_altam {at} yahoo {dot} com United States
Amitabha Majumdar Mamitava {at} nccs {dot} res {dot} in India
Keoni Gandall koeng101 {at} gmail {dot} com United States
Jennifer Hsieh j.hsieh {at} kurabiotech {dot} com Chile
Kenneth Frimpong ken.frimpong {at} thrivusinstitute {dot} edu {dot} gh Ghana
Shibichakravarthy Kannan shibi.kannan {at} gmail {dot} com India
Ganesh Prasad ganeshmayya {at} gmail {dot} com India
Stephen Klusza smklusza {at} gmail {dot} com United States
Long Nguyen Lnguyen2 {at} ufl {dot} edu United States
Ahmed Atef ahmed_atefaig2 {at} yahoo {dot} com Saudi Arabia
Emin Bursa NA United States
raghav sridhar raghav {at} cambrianbioworks {dot} com India
Nadia Odaliz Chamana Chura nadia.chamana {at} utec {dot} edu {dot} pe France
Benjamin Loang benjamin.liang {at} uq {dot} edu {dot} au Australia
Stephen Klusza StephenKlusza {at} clayton {dot} edu United States
Tomasz Jurkowski jurkowskit {at} cardiff {dot} ac {dot} uk United Kingdom
Marko Komlosh markokomlosh {at} gmail {dot} com Serbia
Majed Alghamdi mafa2 {at} le {dot} ac {dot} uk United Kingdom
Wei-Min Chang WeiMinChang {at} tmu {dot} edu {dot} tw Taiwan
Dong Nguyen Tam NA Vietnam
Zhiyong Lei zlei {at} umcutrecht {dot} nl Netherlands
Tri Ngo NA United States
Kefentse Arnold Tumedi tumediak {at} yahoo {dot} com Botswana
Maira Goytia mgoytia {at} spelman {dot} edu United States
Amjed Alsultan amjed.talib {at} qu {dot} edu {dot} iq Iraq
Norhan Hassan norhan.hassan {at} kaust {dot} edu {dot} sa Saudi Arabia
Jose David Rosales https://www.linkedin.com/in/david-rosales-574a7151/ United States
Louise Dalgaard ltd {at} ruc {dot} dk Denmark
Esteban Erben eerben {at} iib {dot} unsam {dot} edu {dot} ar Argentina
Jan Studeny jan.studeny {at} aalto {dot} fi Finland
Vincent Paris NA United States
Jimmy Gledson Hayden Linhares haydenlinhars {at} gmail {dot} com Brazil
Johan Sosa johan {at} thefunquantum {dot} com United States
Ben Davis bed58 {at} aber {dot} ac {dot} uk United Kingdom
Rodolfo Marcolino rodolfo.marcolino {at} me {dot} com Argentina
Matthew Phanchana matthew.pha {at} mahidol {dot} edu Thailand
Jordan Gonzalez jgonzalez {at} thecitizensciencelab {dot} org United States
Ingrid Lorena Jaramillo Lojadel2 {at} gmail {dot} com Colombia
Krishna Gupta gupta-krishna {at} ipfdd {dot} de Germany
Ahmad Suparmin ahmad.suparmin {at} ugm {dot} ac {dot} id Indonesia
TING-WEI LIN weitinglin66 {at} gmail {dot} com Taiwan
Deepak Saini deepaksaini {at} iisc {dot} ac {dot} in India
Joey Riepsaame joey.riepsaame {at} path {dot} ox {dot} ac {dot} uk United Kingdom
Jose David Rosales jdrr55 {at} yahoo {dot} com https://www {dot} linkedin {dot} com/in/david-rosales-574a7151/ Venezuela
Pratik Vyas iampratikvyas {at} gamil {dot} com India
Han Teng Wong wong_han_teng {at} imcb {dot} a-star {dot} edu {dot} sg Singapore
Bingbing Wan wanb {at} sjtu {dot} edu {dot} cn China
Leandro Oliveira leandro.licursi {at} ufv {dot} br United Kingdom
Xavier Coadic xcoadic {at} protonmail {dot} com France
Simran Kushwaha https://www.bits-pilani.ac.in/pilani/biologicalScience/Faculty India
Irfan Hussain irfan {at} bs {dot} qau {dot} edu {dot} pk Pakistan
Joao Vitor Dutra Molino candidomolino {at} gmail {dot} com United States
Vitaliy Strochkov NA Kazakhstan
Andrei Berasneu andreyberesnevson {at} gmail {dot} com Poland
Ahmed Hegazy a.hegazy {at} biotec {dot} rwth-aachen {dot} de Germany
Codruta Ignea codruta.ignea {at} mcgill {dot} ca Canada
Ahmed Atef ahmed_atefaig2 {at} yahoo {dot} com United States
Carlos Ueira-Vieira ueira {at} ufu {dot} br Brazil
Cihan Aydin cihan.aydin {at} medeniyet {dot} edu {dot} tr Turkey

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