Simultaneous DNA Typing of Human Platelet Antigens 2, 3 and 4 by an Allele-Specific PCR Method

Vox Sanguinis ◽  
1995 ◽  
Vol 68 (4) ◽  
pp. 225-230
Author(s):  
Shigenori Tanaka ◽  
Atsuko Taniue ◽  
Nobuo Nagao ◽  
Shiro Ohnoki ◽  
Hirotoshi Shibata ◽  
...  
Keyword(s):  
Human Platelet ◽  
Dna Typing ◽  
Specific Pcr ◽  
Allele Specific ◽  
Pcr Method ◽  
Allele Specific Pcr

Simultaneous DNA Typing of Human Platelet Antigens 2, 3 and 4 by an Allele-Specific PCR Method

Vox Sanguinis ◽  
1995 ◽  
Vol 68 (4) ◽  
pp. 225-230 ◽  
Author(s):  
Shigenori Tanaka ◽  
Atsuko Taniue ◽  
Nobuo Nagao ◽  
Shiro Ohnoki ◽  
Hirotoshi Shibata ◽  
...  
Keyword(s):  
Human Platelet ◽  
Dna Typing ◽  
Specific Pcr ◽  
Allele Specific ◽  
Pcr Method ◽  
Allele Specific Pcr

scholarly journals Use of site-directed mutagenesis of allele-specific PCR primers to identify the GSTM1 A, GSTM1 B, GSTM1 A,B and GSTM1 null polymorphisms at the glutathione S-transferase, GSTM1 locus

1993 ◽  
Vol 295 (1) ◽  
pp. 313-315 ◽  
Author(s):  
A A Fryer ◽  
L Zhao ◽  
J Alldersea ◽  
W R Pearson ◽  
R C Strange
Keyword(s):  
Pcr Primers ◽  
Single Step ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Glutathione S Transferase ◽  
Specific Pcr ◽  
Target Dna ◽  
Allele Specific ◽  
Pcr Method ◽  
Allele Specific Pcr

We describe the identification of the GSTM1 null, GSTM1 A, GSTM1 B and GSTM1 A,B polymorphisms at the glutathione S-transferase GSTM1 locus using a single-step PCR method. Target DNA was amplified using primers to intron 6 and exon 7 with site-directed mutagenesis being used to introduce a restriction site in DNA amplified from GSTM1 *A, thereby allowing differentiation of this allele and GSTM1 *B. The accuracy of this approach in identifying the GSTM1 A, GSTM1 B, GSTM1 A,B and GSTM1 null polymorphisms was confirmed by comparison with, firstly, an established PCR method that distinguishes GSTM1 *0 homozygotes from individuals with the other GSTM1 genotypes and, secondly, GSTM1 phenotypes determined using chromatofocusing.

scholarly journals Screening for deafness-associated mitochondrial 12S rRNA mutations by using a multiplex allele-specific PCR method

2020 ◽  
Vol 40 (5) ◽  
Author(s):  
Yu Ding ◽  
Jianyong Lang ◽  
Junkun Zhang ◽  
Jianfeng Xu ◽  
Xiaojiang Lin ◽  
...  
Keyword(s):  
Hearing Loss ◽  
Age At Onset ◽  
12S Rrna ◽  
Specific Pcr ◽  
Functional Variants ◽  
Sanger Sequence ◽  
Allele Specific ◽  
Mitochondrial 12S Rrna ◽  
Pcr Method ◽  
Allele Specific Pcr

Abstract Mitochondrial 12S rRNA A1555G and C1494T mutations are the major contributors to hearing loss. As patients with these mutations are sensitive to aminoglycosides, mutational screening for 12S rRNA is therefore recommended before the use of aminoglycosides. Most recently, we developed a novel multiplex allele-specific PCR (MAS-PCR) that can be used for detecting A1555G and C1494T mutations. In the present study, we employed this MAS-PCR to screen the 12S rRNA mutations in 500 deaf patients and 300 controls from 5 community hospitals. After PCR and electrophoresis, two patients with A1555G and one patient with C1494T were identified, this was consistent with Sanger sequence results. We further traced the origin of three Chinese pedigrees. Clinical evaluation revealed variable phenotypes of hearing loss including severity, age at onset and audiometric configuration in these patients. Sequence analysis of the mitochondrial genomes from matrilineal relatives suggested the presence of three evolutionarily conserved mutations: tRNACys T5802C, tRNALys A8343G and tRNAThr G15930A, which may result the failure in tRNAs metabolism and lead to mitochondrial dysfunction that was responsible for deafness. However, the lack of any functional variants in GJB2, GJB3, GJB6 and TRMU suggested that nuclear genes may not play active roles in deafness expression. Hence, aminoglycosides and mitochondrial genetic background may contribute to the clinical expression of A1555G/C1494T-induced deafness. Our data indicated that the MAS-PCR was a fast, convenience method for screening the 12S rRNA mutations, which was useful for early detection and prevention of mitochondrial deafness.

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