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Unformatted text preview: the Jatoi et al. (2006) study of the family
Zingiberaceae. The primers were RM1, RM117, RM125,
RM131, RM135, RM153, RM154, and RM171.
PCR amplification was performed twice for each
primer to ensure their reproducibility. The Biometra T1
Thermocycler (Biometra GmbH, Goettingen, Germany)
was used for PCR amplification reactions. The final
volume of the reaction mixture that was used in the
PCR analysis was 25 µL. The solution contains 12.5 µL of
DreamTaq Green PCR Master Mix (2X) (Fermentas, First
Base Laboratories Ptd. Ltd., Seri Kembangan, Selangor,
Malaysia), 0.5 mM of MgCl2 (Fermentas), 0.4 μm of primer
(Fermentas), and 50 ng of DNA template. The thermal
profile of the reaction was programmed as follows: initial
denaturation at 94 °C for 5 min, followed by 35 cycles at 94
°C for 1 min, annealing at 60 °C for 1 min, and extension
at 72 °C for 1 min and 30 s. The final extension was at 72 °C
for 10 min followed by cooling to 4 °C. PCR products were
separated by electrophoresis through 1.2% agarose gels in
1X TAE buffer at 70 V for 75 min. The gels were stained in ethidium bromide...
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- Spring '14