A comprehensive experiment for molecular biology: Determination of single nucleotide polymorphism in human REV3 gene using PCR-RFLP

Biochem Mol Biol Educ. 2017 Jul 8;45(4):299-304. doi: 10.1002/bmb.21037. Epub 2017 Feb 1.

Abstract

Laboratory exercise is helpful for medical students to understand the basic principles of molecular biology and to learn about the practical applications of molecular biology. We have designed a lab course on molecular biology about the determination of single nucleotide polymorphism (SNP) in human REV3 gene, the product of which is a subunit of DNA polymerase ζ and SNPs in this gene are associated with altered susceptibility to cancer. This newly designed experiment is composed of three parts, including genomic DNA extraction, gene amplification by PCR, and genotyping by RFLP. By combining these activities, the students are not only able to learn a series of biotechniques in molecular biology, but also acquire the ability to link the learned knowledge with practical applications. This comprehensive experiment will help the medical students improve the conceptual understanding of SNP and the technical understanding of SNP detection. © 2017 by The International Union of Biochemistry and Molecular Biology, 45(4):299-304, 2017.

Keywords: Laboratory exercise; PCR-RFLP; SNP; molecular biology.

MeSH terms

  • DNA / genetics
  • DNA / isolation & purification
  • DNA / metabolism
  • DNA Primers / chemistry
  • DNA Primers / metabolism*
  • DNA-Binding Proteins / genetics*
  • DNA-Directed DNA Polymerase / genetics*
  • Educational Measurement / statistics & numerical data*
  • Electrophoresis, Agar Gel
  • Gene Expression
  • Humans
  • Leukocytes, Mononuclear / chemistry
  • Leukocytes, Mononuclear / metabolism
  • Molecular Biology / education*
  • Polymerase Chain Reaction / methods
  • Polymorphism, Restriction Fragment Length*
  • Polymorphism, Single Nucleotide*
  • Students

Substances

  • DNA Primers
  • DNA-Binding Proteins
  • DNA
  • DNA-Directed DNA Polymerase
  • REV3L protein, human