A new thermal cycling mechanism for effective polymerase chain reaction in microliter volumes
Journal
Microsystem Technologies
Journal Volume
10
Journal Issue
8-9
Pages
579-584
Date Issued
2004
Author(s)
Abstract
This study presents a new thermal cycler using infrared (IR) heating and water impingement cooling for polymerase chain reaction (PCR) amplification of 10 μl samples in thin glass capillary tubes. The thermal cycling system can achieve a temperature ramping-up rate of 65°C/s and a ramping-down rate of 80°C/s. Two other cooling mechanisms, natural convection and forced air convection, can also be used in the present system to obtain a ramping-down rate of 2°C/s and 6°C/s, respectively. The amplification of the 439 fragment of hepatitis B virus (HBV) DNA was successful. The PCR amplified products were analyzed by agarose gel electrophoresis with ethidium bromide staining for visualization. A comparison of results of the amplification products at three different ramping-down rates was made and the rapid thermal cycling of the present system can run DNA required amplification in 29 min for 20 thermal cycles that is only 1/3 the time spent in the conventional PCR machine used in comparison.
SDGs
Other Subjects
Amplification; Cooling; Enzymes; Infrared heating; Natural convection; Thermal cycling; Visualization; Water; Convetional thermal cycler; Microcentrifuge tubes; Polymerase chain reactions (PCR); DNA
Publisher
Springer Verlag
Type
journal article