Genomic DNA was extracted from whole blood using a standardized salting-out method [18 (link)]. SNP rs6265 (Val66Met) was genotyped using the TaqMan allelic discrimination assay from Applied Biosystems Inc. The predesigned SNP genotyping assay ID is ID C_11592758_10. Real-time PCR was performed in 20 μl volumes with 5 ng of genomic DNA, 1 μl TaqMan SNP genotyping assay (containing two PCR primers and two dye (VIC or FAM)-labeled TaqMan MGB probes) and 10 μl of TaqMan Universal PCR Master Mix (Applied Biosystem) according to the manufacturer’s manual. PCR was performed at 95 °C for 10 min and 40 cycles at 95 °C for 15 s and 60 °C for 1 min using the ABI Prism 7900HT RealTime PCR (Applied Biosystems). Analyzes of amplification products were achieved using SDS software, version 2.4. Two blank controls in each 96-well-plate were used for the assay quality control. Subjects were genotyped as follows: homozygous for the Val allele (ValVal), heterozygotes (Val-Met), and homozygous for the Met allele (MetMet).
Abi prism 7900ht realtime pcr
Manufactured by Thermo Fisher Scientific
The ABI Prism 7900HT Real-Time PCR System is a laboratory instrument designed for real-time polymerase chain reaction (PCR) analysis. It is capable of performing quantitative gene expression analysis and genetic variation detection.
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Lab products found in correlation
Taqman allelic discrimination assay, by Thermo Fisher Scientific (1 mentions)
Taqman universal pcr master mix, by Thermo Fisher Scientific (1 mentions)
High capacity cdna archive kit, by Thermo Fisher Scientific (1 mentions)
Taqman fam labeled specific probes, by Thermo Fisher Scientific (1 mentions)
2 protocols using abi prism 7900ht realtime pcr
1
Genotyping BDNF Val66Met Polymorphism
2
Random-primed cDNA Synthesis and RT-qPCR
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Random-primed cDNA synthesis was performed at 37°C starting with 0.3 μ g of RNA, using the High Capacity cDNA Archive kit (Applied Biosystems). Gene expression was measured in an ABI Prism 7900HT Real Time PCR system using TaqMan FAM-labeled specific probes (Applied Biosystems). A list of the probes used is presented in Supplementary Table 1 (S1 ). Results were normalized to TATA-binding protein (Tbp) expression.
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