Study of the Modification of Loop-mediated Isothermal Amplification (LAMP) using Taq Polymerase for Halal Testing

Rosy Hutami, Mira Suprayatmi, Nida Idzni, Raafqi Ranasasmita, Henny Nuraini, Joko Hermanianto

2018

Abstract

Loop-Mediated Isothermal Amplification (LAMP) is a new method in nucleic acid analysis that amplifies target in isothermal or constant conditions. This method is suitable for analysis with low-resource condition. Commonly, this method needs a polymerase enzyme that has strand displacement activity such as Bacillus stearothermopilus (Bst) polymerase. But, this enzyme was rarely available in the laboratory. Taq polymerase is an polymerase enzyme that usualy available in molecular laboratory, easier to be accessed, and has a lower cost comparing to the Bst polymerase. This research conducted with Taq polymerase in order to study the compatibility of taq polymerase in amplificating the deoxyribonucleic acid (DNA) with LAMP method. We used cytochrome b (cyt b) from pork as terget gene for halal detection need. For conducting the strand displacement activity in DNA amplification, we used denaturation process (95oC), following by annealing (65oC), and enzyme inactivation (80oC). The result showed there was no band appeared on the agarose after electrophoresis. It was suggested that the Taq polymerase was not suitable for LAMP analysis although it has been combined with denaturation process. The LAMP should be conducted with a suitable polymerase that has strand displacement activity and some supporting reagents such as betaine and LAMP buffer that can strengthen the reaction under isothermal condition.

Paper Citation

in Harvard Style

Hutami R., Suprayatmi M., Idzni N., Ranasasmita R., Nuraini H. and Hermanianto J. (2018). Study of the Modification of Loop-mediated Isothermal Amplification (LAMP) using Taq Polymerase for Halal Testing.In Proceedings of the 1st International Conference on Recent Innovations - Volume 1: ICRI, ISBN 978-989-758-458-9, pages 2191-2198. DOI: 10.5220/0009940721912198

in Bibtex Style

@conference{icri18,
author={Rosy Hutami and Mira Suprayatmi and Nida Idzni and Raafqi Ranasasmita and Henny Nuraini and Joko Hermanianto},
title={Study of the Modification of Loop-mediated Isothermal Amplification (LAMP) using Taq Polymerase for Halal Testing},
booktitle={Proceedings of the 1st International Conference on Recent Innovations - Volume 1: ICRI,},
year={2018},
pages={2191-2198},
publisher={SciTePress},
organization={INSTICC},
doi={10.5220/0009940721912198},
isbn={978-989-758-458-9},
}

in EndNote Style

TY - CONF

JO - Proceedings of the 1st International Conference on Recent Innovations - Volume 1: ICRI,
TI - Study of the Modification of Loop-mediated Isothermal Amplification (LAMP) using Taq Polymerase for Halal Testing
SN - 978-989-758-458-9
AU - Hutami R.
AU - Suprayatmi M.
AU - Idzni N.
AU - Ranasasmita R.
AU - Nuraini H.
AU - Hermanianto J.
PY - 2018
SP - 2191
EP - 2198
DO - 10.5220/0009940721912198