Molecular Diagnostics Toolkit

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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	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

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
Ahmed Hegazy	a.hegazy {at} biotec {dot} rwth-aachen {dot} de
TING-WEI LIN	weitinglin66 {at} gmail {dot} com
raghav sridhar	raghav {at} cambrianbioworks {dot} com
Vitaliy Strochkov	NA
Amitabha Majumdar	Mamitava {at} nccs {dot} res {dot} in
Jordan Gonzalez	jgonzalez {at} thecitizensciencelab {dot} org
Kenneth Frimpong	ken.frimpong {at} thrivusinstitute {dot} edu {dot} gh
Matthew Phanchana	matthew.pha {at} mahidol {dot} edu
Amjed Alsultan	amjed.talib {at} qu {dot} edu {dot} iq
Jimmy Gledson Hayden Linhares	haydenlinhars {at} gmail {dot} com
Keoni Gandall	koeng101 {at} gmail {dot} com
Louise Dalgaard	ltd {at} ruc {dot} dk
James Altamirano	James_altam {at} yahoo {dot} com
Ahmed Atef	ahmed_atefaig2 {at} yahoo {dot} com
Dong Nguyen Tam	NA

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MTA

Unilateral OpenMTA

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