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antispasmodic and sedative [13-16]. Three new sesquiterpene. Genomic DNA was isolated from peripheral blood leukocytes using the phenol-chloroform method [15, 17]. The ZNF259 rs2075290 SNP was genotyped by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). PCR amplification was performed using 5'-GAGTCTATGGGATAATGTGC-3' as the forward and 5'-AATTCCAGGCCTAACTTTGG-3' as reversed primer pair. Each amplification reaction was performed in a total volume of 25 ml, 12.5 μl of 2 × Taq PCR MasterMix (constituent: 0.1 U Taq polymerase/μl, 500 μM dNTP each and PCR buffer) and nuclease-free water 8.5 μl, 20 pmol/L of each primer and 100 ng of genomic DNA, processing started with 5 min of pre-denaturing at 95 °C and followed by 45 s of denaturing at 94 °C, 30 s of annealing at 56.7 °C and 1 min of elongation at 72 °C for 35 cycles. The amplification was completed by a final extension at 72 °C for 10 min. Then each restriction enzyme reaction was performed with 10 μl of amplified DNA, 8 μl of nuclease-free water, 1 μl of 10 × buffer solution, and 10 U of SspI enzyme in a total volume of 20 μl digested at 37°C overnight. After restriction enzyme digestion of the amplified DNA, the digestive products were separated by electrophoresis on 2% agrose gel. The length of each digested DNA fragment was determined by comparing migration of a sample with that of standard DNA marker. Genotypes were scored by an experienced reader blinded to the epidemiological and lipid results. Six samples (each genotype in two; respectively) detected by the PCR-RFLP were also confirmed by direct sequencing. The PCR products were purified by low melting point gel electrophoresis and phenol extraction, and then the DNA sequences were analyzed using an ABI Prism 3100 (Applied Biosystems) in Shanghai Sangon Biological Engineering Technology & Services Co., Ltd., People's Republic of China.. there are many different names for
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Unani and others are categorized under alternative. signal. Multicellular organisms utilize a set of Notch pathway receptors. mitochondrial protein synthesis, growing cells were labeled with [35S]. pelvic floor.
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C4-II, to killing by wt and M protein mutant VSV strains, cells were.
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