Genetic Analysis of CYP27A1 Gene

Genomic DNA was extracted from peripheral blood samples using a commercial blood genomic extraction kit (Qiagen, Hilden, Germany). Polymerase chain reaction (PCR) was carried out to amplify all exons and flanking regions of CYP27A1. Direct Sanger sequencing was performed on an ABI 3500xl Dx Genetic Analyzer (Applied Biosystems, Foster City, CA, USA) as described previously [28 (link)]. The primers for CYP27A1 were listed in Additional file 1: Table S1. The 1000 Genomes Project (https://www.ncbi.nlm.nih. gov/variation/tools/1000 genomes/) and the ExAC database (https://exac.broadinstitute.org/) were used to check the frequency of variants in the general population. Three software programs, including SIFT (http://sift.jcvi.org/), PolyPhen-2 (http://genetics.bwh.harvard.edu/pph2/) and Mutation Taster (http://www.mutationtaster.org/) were used to predict the possible protein functional changes caused by the variants.

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Tao Q.Q., Zhang Y., Lin H.X., Dong H.L., Ni W, & Wu Z.Y. (2019). Clinical and genetic characteristics of Chinese patients with cerebrotendinous xanthomatosis. Orphanet Journal of Rare Diseases, 14, 282.

Publication 2019

blood genomic extraction kit ABI 3500xl Dx Genetic Analyzer

Blood Exons Genetic Genomes Mutation Polymerase chain reaction Primers Protein changes

Corresponding Organization :

Other organizations : Second Affiliated Hospital of Zhejiang University

Top 5 similar protocols

Variable analysis

independent variables

Genomic DNA extraction from peripheral blood samples using a commercial blood genomic extraction kit (Qiagen, Hilden, Germany)
Polymerase chain reaction (PCR) to amplify all exons and flanking regions of CYP27A1
Direct Sanger sequencing on an ABI 3500xl Dx Genetic Analyzer (Applied Biosystems, Foster City, CA, USA)

dependent variables

Sequence of CYP27A1 gene

control variables

Primers for CYP27A1 listed in Additional file 1: Table S1
Checking the frequency of variants in the general population using the 1000 Genomes Project and the ExAC database
Predicting the possible protein functional changes caused by the variants using SIFT, PolyPhen-2, and Mutation Taster software

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