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July 29, 2024
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Post in: Application Notes, Life Sciences Research
NZYtech’s LAMP and RT-LAMP Positive Control kits: invaluable tools in isothermal test development and use
João S. Teodoro¹ Rita S. Simões¹´² & Carlos M.G.A. Fontes¹
¹ Department of Research and Development, NZYtech, Lisbon, Portugal; [2] CIISA, Faculty of Veterinary Medicine, University of Lisbon, Portugal
* joao.teodoro@nzytech.com
Introduction
Isothermal amplification techniques such as loop-mediated isothermal amplification (LAMP) have gathered recent attention due to their immediate advantages over traditional qPCR testing. LAMP assays do not require thermal cycling, reducing equipment needs and improving time to result values due to the technique’s intricate steps and reaction sequences, as well as the elevated processivity of its critical enzyme, Bst DNA polymerase[1, 2].
Although LAMP assays offer high target specificity, the complex reaction matrix which involves up to six primer sequences and a relatively low reaction temperature (55-65 °C) often leads to non-specific amplifications[3]. Adding to this, primer design requirements are less straightforward than a typical PCR reaction, while probe insertion is a challenging design task due to the lack of 5’à3’ exonuclease activity by Bst[4,5]. These hurdles are manageable in well-established LAMP and RT-LAMP protocols, but in earlier phases of development, in troubleshooting established protocols, where certitude of result is arguably the most critical, these might render progress prohibitively slow and costly.
It is with these issues in mind that NZYtech has developed LAMP and RT-LAMP Positive Control Kits, two novel products that contain all the individualized components required to produce a fully functional LAMP/ RT-LAMP reaction, or to mix and match different reaction components to assess their usefulness. In this matter, these kits allow for a quick, cost-efficient and reliable means to test new reagents, primers, enzymes or any other reaction elements one might want to test. They are also a great tool for any researcher starting out in isothermal amplification procedures, as well as powerful teaching aides, allowing for customizable and hands-on learning of the role and importance of the various elements of a LAMP/ RT-LAMP reaction.
In this application note, we detail and characterize the use of NZYtech’s LAMP and RT-LAMP Positive Control kits, with the suggestion of a reaction protocol and tips for overcoming any issues one might have in isothermal amplification.
Application Note