Genomic DNA was extracted from whole blood using the Autopure LS (Qiagen, Hilden, Germany). All three single-nucleotide polymorphisms (SNP; PNPLA3 at rs738409, C>G/I148M; TM6SF2 at rs58542926, C>T/E167K; and MBOAT7 at rs641738, C>T) were genotyped by TaqMan PCR method (Applied Biosystems, Foster City, CA) according to manufacturer’s instructions. Post-PCR allelic discrimination was carried out measuring allele-specific fluorescence on an ABI Prism Sequence Detection System ABI 7900HT (Applied Biosystems). The success rate for genotyping was >95% for all three SNPs and the genotypes of all three SNPs were in Hardy–Weinberg equilibrium.
Abi prism sequence detection system abi 7900ht
Manufactured by Thermo Fisher Scientific
The ABI Prism Sequence Detection System ABI 7900HT is a real-time PCR system designed for gene expression analysis and other quantitative nucleic acid detection applications. The system utilizes thermal cycling technology to amplify and detect target DNA sequences in real-time. It provides quantitative data on nucleic acid levels within a sample.
Automatically generated - may contain errors
2 protocols using abi prism sequence detection system abi 7900ht
1
Genotyping of Liver Disease SNPs
2
Genotyping of Genetic Variants
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Genomic DNA was extracted from 3 mL of ethylenediaminetetraacetic acid whole blood samples using the salt fractionation method, which was standardized in the Puregene kit commercialized by Gentra (Research Triangle, NC) and was stored at −80°C. Approximately 10 ng of DNA was used for genotyping by TaqMan polymerase chain reaction (Applied Biosystems, Foster City, CA) according to the manufacturer's directions. After thermal cycling (50°C for 2 minutes, 95°C for 10 minutes, and then 40 cycles at 95°C for 15 seconds and at 60°C for 90 seconds), allelic discrimination for rs8192678 (cat.C_1643192_20) in PPARGC1A, rs738409 (cat.C_7241_10) in PNPLA3, and rs780094 (cat.C_2862873_10) in GCKR was performed by measuring the allele-specific fluorescence on an ABI Prism Sequence Detection System ABI 7900HT (Applied Biosystems).
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