COVID-19 is currently destroying hundreds of thousands of lives around the world. Widespread testing for this virus is necessary to control its spread, but in many places, conducting tests costs too much money because of the reagent cost. We have collected expression plasmids for the most expensive test components, the enzymes. With these expression plasmids, users can produce the enzymes needed to do testing locally much cheaper than it would cost to purchase or import them, which we hope will allow for more COVID-19 tests.
What is it:
This is a collection of 5 different protein-coding genes in expression vectors. We have 2 reverse transcriptases (HIV-RT and MMLV-RT), two DNA polymerases (Taq and Bst-LF), and one RNA inhibitor. All vectors, except HIV-RT, are ampicillin resistant, with HIV-RT being kanamycin resistant. The RNAse inhibitor plasmid and the Taq polymerase plasmid both use the tac promoter, while the HIV-RT, MMLV-RT, and Bst-LF plasmids use the T7 promoter.
Unlike most of the collections that FreeGenes offers, all 5 expression vectors will be shipped as raw DNA. You do not need a -80C freezer to store this kit. If you are asked at checkout whether you possess a -80C freezer, you can still receive this kit if you answer 'no.'
What can it be used for:
These enzymes can generally be used for reverse transcription and PCR or LAMP amplification. Any protocol that requires any of those 3 reactions can use the enzymes expressed in these vectors.
These materials were created by the Dickinson lab at UC Chicago, Alex Brown at National Jewish Health, John Schloendorn at Gene And Cell Technology, and the Open Bioeconomy Lab at Cambridge University.
Where I can find more information:
- pOpenTaq Protocol
- HIV-RT Protocol
- MMLV-RT Protocol
- Bst-LF Protocol
- Open Bioeconomy Lab (for troubleshooting MMLV-RT and Bst-LF)
- Certification Data
Here are the genbank format files for each detection enzyme expression vector. You will need the free SnapGene® viewer or other program.
- Genbank file for HIV-RT expression plasmid
- Genbank file for RNAse Inhibitor expression plasmid
- Genbank file for Bst-LF expression plasmid
- Genbank file for MMLV-RT expression plasmid
- Genbank file for Taq expression plasmid
This product is made available under the unilateral OpenMTA.
The items in this collection can also be obtained from Addgene as individual items:
- Taq polymerase
- RNAse Inhibitor
- Bst Large Fragment
- MMLV Reverse Transcriptase
- HIV Reverse Transcriptase
|HIV-RT||His-tagged HIV-RT p66|
|Bst-LF||Bst-LF DNA polymerase|
|RNAse Inhibitor||RNAse Inhibitor|