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Updated: May 21, 2026

Candidate Gene Testing in Clinical Cohort Studies with Multiplexed Genotyping and Mass Spectrometry
Published on: June 21, 2018
Sequence-Specific Primer Polymerase Chain Reaction Genotyping of Interleukin-22 Single Nucleotide Polymorphisms
Julyane Schavaren1, Matheus Braga1, Aléia Harumi Uchibaba Yamanaka1
1Biosciences and Physiopathology Program, State University of Maringá, Maringá, Brazil.
Abstract:
Single nucleotide polymorphisms (SNPs) can modify protein expression or function, influencing susceptibility to various diseases. Interleukin-22 (IL-22), involved in epithelial barrier homeostasis and tissue repair, has shown complex roles in autoimmune, infectious, and inflammatory conditions. This article describes a standardized protocol for genotyping the IL-22 SNPs rs2227473 (C>T) and rs2227513 (A>G) using sequence-specific primer polymerase chain reaction (PCR-SSP), with validation by Sanger sequencing. The protocol includes optimization of annealing temperature, MgCl2 concentration, primer balance, deoxynucleotide triphosphates (dNTPs), and DNA input, using the human growth hormone (HGH) gene as an internal control. Amplified products are analyzed by agarose gel electrophoresis. Complete concordance with Sanger sequencing confirms the reliability of the method. This protocol provides a rapid, cost-effective, and reproducible approach for SNP genotyping, suitable for large-scale studies. © 2026 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol: PCR-SSP genotyping of IL-22 SNPs rs2227473 and rs2227513 Support Protocol: Validation of genotypes by Sanger sequencing.

