《Table 1 Specimens and sequences of three new species of Vietnamophryne Gen.nov.from Indochina and o

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《A new genus and three new species of miniaturized microhylid frogs from Indochina(Amphibia: Anura:Microhylidae: Asterophryinae)》

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The PCR products were loaded onto 1.5%agarose gels in the presence of ethidium bromide.Visualization was carried out using agarose electrophoresis.If distinct bands were obtained，products were purified prior to cycle sequencing using 2μL of Exo Sap It（Amersham，UK），diluted at a 1:4ratio，per 5μL of PCR product.The 10μL sequencing reaction included 2μL of template，2.5μL of sequencing buffer，0.8μL of 10 pmol primer，0.4μL of Big Dye Terminator v3.1Sequencing Standard（Applied Biosystems，USA），and 4.2μL of water.The cycle sequencing reaction included 35 cycles with the following steps:10 s at 96?C，10 s at 50?C，and 4min at 60?C.Cycle sequencing products were then purified by ethanol precipitation.Sequencing was performed on an ABI 3730xl automated sequencer（Applied Biosystems，USA）.The obtained sequences were deposited in Gen Bank under accession numbers MH004403–MH004406（Table 1）.