Dna quantities and qualities from various stages of some trematodes using optical and HAT-RAPD methods.
Southeast Asian J Trop Med Public Health
; 2006 ; 37 Suppl 3(): 62-8
Article
in En
| IMSEAR
| ID: sea-32988
ABSTRACT
The aim of this experiment was to minimize DNA quantity and quality for detection by optical (spectrophotometer at 260 nm and 280 nm) and HAT-RAPD methods. Total DNA from different stages, adult, metacercaria and eggs of 6 trematode species were isolated for analysis. In this experiment, the adult trematodes were classified into 3 groups by size small, Haplorchis taichui and Stellantchasmus falcatus; medium, Opisthorchis viverrini and Ganeo tigrinus; and large, Paramphistomum epiclitum and Fischoederius elongatus. The adult minimal DNA quantities and qualities of all specimen samples detected by optical method were 97.22,72.28, 3,167.00, 1,490.62, 21,382.66, and 27,321.77 ng; eggs were 3.92, 3.57, 3.72, 6.23, 17.53, and 14.01 ng, respectively; and metacercarial stages 50.70 and 40.98 ng in H. taichui and S. falcatus. In addition, the HAT-RAPD technique was chosen to amplify the minimal DNA qualities and quantities of all trematode specimens. Total DNA was 1-1 x 10(-12) ng; DNA templates in each dilution were used for amplification by primer OPA-09. DNA concentrations ranging between 1 x 10(-8) and 1 x 10(-11) ng were amplified with high polymorphism. Our experiment concluded that only a single specimen of each egg, metacercaria, or adult stage could be amplified with distinct bands.
Full text:
1
Index:
IMSEAR
Main subject:
Trematoda
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Polymerase Chain Reaction
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DNA, Helminth
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Random Amplified Polymorphic DNA Technique
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Electrophoresis, Agar Gel
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Life Cycle Stages
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Animals
Language:
En
Journal:
Southeast Asian J Trop Med Public Health
Year:
2006
Type:
Article