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A laboratory exercise to determine human ABO blood type by noninvasive methods
Author(s) -
Martin Michael P.,
Detzel Stephen M.
Publication year - 2008
Publication title -
biochemistry and molecular biology education
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.34
H-Index - 39
eISSN - 1539-3429
pISSN - 1470-8175
DOI - 10.1002/bmb.20165
Subject(s) - abo blood group system , genomic dna , biology , genetics , exon , allele , microbiology and biotechnology , agarose gel electrophoresis , saliva , buccal swab , dna , gene , biochemistry
Analysis of single‐nucleotide polymorphisms and their association with diseases and nondisease phenotypes is of growing importance in human biology studies. In this laboratory exercise, students determine the genetic basis for their ABO blood type; however, no blood is drawn. Students isolate genomic DNA from buccal mucosa cells that are present in saliva and analyze the DNA on an agarose gel. Subsequently, this DNA is used as a PCR template to amplify exons 6 and 7 of the gene that determines the human ABO phenotype. These PCR products are digested and run on agarose gels to examine the restriction fragment length polymorphisms. A deletion in the O 1 allele converts the Bst E II site in exon 6 into a Kpn I site, and this feature is used to determine the presence of O 1 alleles. The pattern of exon 7 digest products allows students to distinguish among four other common ABO alleles: A 1 , A 2 , B , and O 2 . This exercise introduces students to commonly used molecular biology techniques, such as genomic DNA isolation, PCR, gel electrophoresis, and restriction digests.