Sanger sequencing was performed on an ABI-A3500 genetic analyzer, and a BigDye 3.1 sequencing kit (Applied Biosystems, California, USA) was used. The primers used for PCR and Sanger sequencing are provided in online supplementary table S1 .
Abi a3500 genetic analyzer
Manufactured by Thermo Fisher Scientific
Sourced in United States
The ABI-A3500 genetic analyzer is a laboratory instrument used for DNA sequencing and genetic analysis. It utilizes capillary electrophoresis technology to separate and detect fluorescently labeled DNA fragments. The ABI-A3500 is designed to provide accurate and reliable data for a variety of genetic applications.
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Lab products found in correlation
2 protocols using abi a3500 genetic analyzer
1
Sanger Sequencing Protocol for Genetic Analysis
2
AGXT Gene Mutation Screening and Validation
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For mutation screening, the coding and intron-exon boundary regions of the AGXT gene (NM_000030, according to GRCh37/hg19) were amplified by PCR, using the primers synthesized by a local biotech company (BGI; Shenzhen, China) and designed with Primer32 . The sequencing reaction (BigDye 3.1 Kit, Applied Biosystems, Waltham, MA, United States) of the purified PCR products was performed according to the recommended procedure. The labeled PCR fragments were purified through 70% alcohol precipitation and electrophoresed on an ABI-A3500 genetic analyzer (Applied Biosystems, Waltham, MA, United States). The primer sequences, PCR conditions, and sequencing are shown in Supplementary Table S2 .
For the validation of the AGXT-exon6 mutation (c.667A > C: p.Ser223Arg), two pairs of primers were used as presented inSupplementary Figure S3 .
For the validation of the AGXT-exon6 mutation (c.667A > C: p.Ser223Arg), two pairs of primers were used as presented in
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