Polymerase chain reaction thermal cycling using the programmed tilt displacements of capillary tubes

Hassan Ali Abid, Eric Shen Lin, Jian Wern Ong, Tristan Minifie, Zhixiong Song, Oi Wah Liew, Tuck Wah Ng

Research output: Contribution to journalArticleResearchpeer-review

Abstract

A thermal cycling method, whereby capillary tubes holding polymerase chain reactions are subjected to programmed tilt displacements so that they are moved using gravity over three spatial regions (I, II, and III) kept at different constant temperatures to facilitate deoxyribonucleic acid (DNA) denaturation, annealing, and extension, is described. At tilt speeds in excess of 0.2 rad/s, the standard deviation of static coefficient of friction values was below 0.03, indicating in sync movement of multiple capillary tubes over the holding platform. The travel time during the acceleration phase and under constant velocity between adjacent regions (I to II and II to III) and distant regions (III to I) was 0.03 s and 0.31 s, respectively. The deviations in temperature did not exceed 0.05 °C from the average at the prescribed denaturing, annealing, and extension temperatures applied. DNA amplification was determined by optical readings, the fluorescence signal was found to increase twofold after 30 thermal cycles, and 1.16 × 106 DNA copies/μl could be detected. The approach also overcomes problems associated with thermal inertia, sample adhesion, sample blockage, and handling of the reaction vessels encountered in the other thermal cycling schemes used.

Original languageEnglish
Article number104105
Number of pages8
JournalReview of Scientific Instruments
Volume91
Issue number10
DOIs
Publication statusPublished - Oct 2020

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