Open AccessLigase chain reaction assay for human mutations: the Sickle Cell by LCR assay
Author(s) -
Antonio A. Reyes,
Paola Carrera,
Elena Cardillo,
Luis Ugozzoli,
Jimmie D. Lowery,
Ching-I P Lin,
Matthew C. Go,
Maurizio Ferrari,
R. Bruce Wallace
Publication year - 1997
Publication title -
clinical chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.705
H-Index - 218
eISSN - 1530-8561
pISSN - 0009-9147
DOI - 10.1093/clinchem/43.1.40
Subject(s) - ligase chain reaction , biotinylation , microbiology and biotechnology , oligonucleotide , polymerase chain reaction , ligation , dna ligase , biology , streptavidin , dna , chemistry , genomic dna , gene , biochemistry , biotin , multiplex polymerase chain reaction
We can detect the beta-globin gene sickle cell mutation by using an assay based on the ligase chain reaction. The simultaneous amplification of the human growth hormone gene in the same reaction serves as a control for the amount of template DNA or amplification efficiency. Ligation products, which are biotinylated at one end and tagged with an arbitrary "tail" sequence at the other, are captured by hybridization to "tail"-complementary oligonucleotides immobilized on polystyrene microwells. The captured ligation products are detected colorimetrically by use of streptavidin-alkaline phosphatase conjugate. In a study of 24 subjects, the assay unequivocally discriminated among normal, carrier, and sickle cell genotypes.
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