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Thermo Scientific™ EquiPhi29™ DNA Amplification Kit
Description
The Thermo Scientific EquiPhi29 DNA Amplification Kit includes all required components for fast, sensitive, and efficient isothermal DNA amplification, which includes rolling circle amplification (RCA) and multiple displacement amplification – whole genome amplification (MDA-WGA).
Features of the EquiPhi29 DNA Amplification Kit include:
• Fast DNA amplification in less than two hours
• Sensitivity from 1 fg of input DNA
• Maximal yield of 17 µg amplified DNA
• Compatibility with different types of sample input material (e.g., purified DNA, liquid media culture, agar plate colonies)
• Compatibility with various downstream applications such as DNA sequencing (Sanger sequencing, next-generation sequencing), digestion with restriction enzymes, cell-free DNA enrichment, and cell-free protein expression
• All required components for DNA amplification in RCA or MDA-WGA reactions
The kit contains EquPhi29 DNA Polymerase, a proprietary phi29 DNA polymerase mutant that possesses strong strand displacement activity and enables high thermostability, reaction speed, and product yield and low amplification bias. Also included is an optimized reaction buffer, exo-resistant random primer, pyrophosphatase, and dNTPs. The addition of pyrophosphatase to the reaction mixture enhances DNA amplification and facilitates sample handling.
Specifications
Specifications
| Product Type | DNA Amplification Kit |
| Form | Liquid |
| For Use With (Application) | RCA, MDA-WGA |
| Polymerase | EquiPhi29 DNA Polymerase |
| Quantity | 500 Reactions |
| Research Category | Nucleic Acid Analysis, In Vitro Transcription, Cell-free Protein Expression, In Situ Sequencing, SNP Detection |
| Content And Storage | • 5 x 100 μL EquiPhi29 DNA Polymerase (10 U/μL) |
| Format | Kit |
| Optimal Reaction Temperature | 42°C |
| Shipping Condition | Dry Ice |
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Frequently Asked Questions (FAQs)
The error rate of EquiPhi29 DNA Polymerase is 6 x 10-6.
The error rate of EquiPhi29 DNA Polymerase was measured according to the method described in literature:
Mielinis, P., Sukackaitė, R., Serapinaitė, A., Samoilovas, F., Alzbutas, G., Matjošaitis, K., & Lubys, A. (2021). MUA-based molecular indexing for rare mutation detection by Next-Generation sequencing. Journal of Molecular Biology, 433(19), 167209. https://doi.org/10.1016/j.jmb.2021.167209
Thermo Scientific EquiPhi29 DNA Polymerase is a proprietary mutant phi29 DNA Polymerase developed through in vitro protein evolution. EquiPhi29 DNA Polymerase as well as phi29 DNA Polymerase should be able to incorporate 5-methyl-dCTP nucleotides and other modified nucleotides.
Cleaning of the amplified product is not required prior to several downstream methods (e.g., debranching, digestion with restriction endonucleases, Sanger sequencing); the dilution of amplified product is sufficient. If the clean-up procedure is needed, we recommend using an affinity-based spin-column or magnetic bead-based purification method.
The optimal reaction time for DNA amplification with EquiPhi29 DNA Polymerase is 2 hours. For samples with ≥1 pg of DNA input, DNA amplification time can be shortened to 1 hour if maximizing product yield is not essential.
Yes. EquiPhi29 DNA Polymerase can work with different types of sample input material such as purified DNA, liquid media culture, agar plate colonies, etc.
For Research Use Only. Not for use in diagnostic procedures.