All the participants underwent DNA sequencing and electrophoretic condition for screening the candidate gene. PCR was performed with PE9700 thermocyclers (Applied Biosystems, Foster City, CA, USA). Sequence analysis was performed on an automated sequencer (ABI 3730, Applied Biosystems) for both affected and normal individuals. Nucleotide alterations including mutations and polymorphisms were identified by sequence alignment with the NCBI (National Center for Biotechnology Information) Reference Sequence using the DNAStar software 5.0 versions (DNASTAR, Madison, WI, USA).
Pe9700 thermocyclers
Manufactured by Thermo Fisher Scientific
Sourced in United States
The PE9700 thermocycler is a laboratory instrument used for the amplification of DNA or RNA samples. It precisely controls the temperature of samples during the thermal cycling process, which is a fundamental step in various molecular biology techniques such as polymerase chain reaction (PCR).
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Lab products found in correlation
3 protocols using pe9700 thermocyclers
1
Genetic Screening Using DNA Sequencing
2
Sanger Sequencing for Rare Deafness Variants
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Sanger sequencing was used to identify participants segregating causative variants in the candidate gene. PCR was performed with a PE9700 thermocyclers (Applied Biosystems, Foster City, CA). Sequence analysis was performed on an automated sequencer (ABI 3730, Applied Biosystems) for both affected and normal individuals. Nucleotide alterations including mutations and polymorphisms were identified by sequence alignment with the NCBI Reference Sequence (NM_001195263.1) using the DNAStar software versions 5.0 (DNASTAR, Madison, WI).
A total of 1,751 normal Chinese individuals served as controls, and 122 Chinese families segregating apparent ARNSHL, who had previously been shown not to have abnormalities in common deafness genes, were ascertained for screening candidate mutations.
A total of 1,751 normal Chinese individuals served as controls, and 122 Chinese families segregating apparent ARNSHL, who had previously been shown not to have abnormalities in common deafness genes, were ascertained for screening candidate mutations.
3
Genetic Variant Analysis of AIFM1
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AIFM1 (NC_000023.10) contains 16 exons. Thirty-two primers (16 primer pairs) were designed using Primer V.3.0 software and synthesised by Invitrogen by Life Technology (Beijing, China) to amplify each exon and exon–intron boundaries (see online supplementary table S1). PCR was performed with PE9700 thermocyclers (Applied Biosystems) using standard conditions. Amplified products from all ANSD cases and controls were gel purified and sequenced (ABI 3730, Applied Biosystems). Nucleotide alterations were identified by sequence alignment with the NCBI Reference Sequence (RefSeq) using DNAStar software V.5.0 (DNASTAR, Madison, Wisconsin, USA).
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