An improved method for MN genotyping by the polymerase chain reaction

Nakayashiki, Nori; Sasaki, Yuki

doi:10.1007/bf01225522

Cited by 11 publications

(8 citation statements)

References 4 publications

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“…In a simple and rapid ASPA method reported by Nakayashiki and Sasaki (1996), conventional M and N atIeles are identified as fragments of different lengths, by shifting the annealing sites of M-and N-specific primers based on three nucleotide substitutions between M and N alleles. Since S and s alleles were attributed to one base change, fragments of the same length were amplified from the alleles in this study.…”

Section: Discussionmentioning

confidence: 99%

“…l) , MN blood group can be genotyped by allele-specific PCR amplification (ASPA) (Corfield et al, 1993;Nakayashiki and Sasaki, 1996) or reverse dot blot hybridization (Herrin et al, 1994); The M-and N-specific sequences are also observed in exon 2 of GPE and GPB genes, respectively ( Fig. 1) (Kudo and Fukuda, 1989Vignal et al, 1990;Rearden et al, 1990;, thus GPA-specific primers should be designed to avoid co-amplification of GPB and GPE segments.…”

Section: Introductionmentioning

confidence: 99%

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PCR-based genotyping of MNSs blood group: Subtyping of M allele to MG and MT

Akane

Kobayashi

et al. 1997

Jap J Human Genet

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SummaryPCR-based genotyping of MNSs blood group system was investigated in combination with restriction fragment length polymorphism (RFLP), single-strand conformation polymorphism (SSCP) and allele-specific PCR amplification (ASPA) techniques. M and N alleles are based on three nucleotide substitutions in exon 2 and one base change (G or T) in an intron of glycophorin A locus. The latter single base change was also found among M alleles analyzed in this study, so that M allele appeared to be subdivided into M c and M T. All three alleles, M c, M r and N were identified clearly by RFLP or SSCP analysis following a single amplification. S and s alleles are based on one nucleotide substitution in exon 3 of glycophorin B gene. Genotyping of Ss blood group system was also explored by PCR-SSCP or ASPA analysis, and problems in the methods were discussed.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

PCR-based genotyping of MNSs blood group: Subtyping of M allele to MG and MT

Akane

Kobayashi

et al. 1997

Jap J Human Genet

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“…MN serological phenotyping and genotyping were performed as reported previously (Nakayashiki and Sasaki 1996;Akane et al 1997;Li et al 1998;Sasaki et al 2000). Two new N alleles had been named temporarily N 2 and N v (Sasaki et al 2000) and are named N201 and N104, respectively, in this paper.…”

Section: Methodsmentioning

confidence: 99%

Systematic classification of alleles of the glycophorin A (MN blood group) gene

et al. 2005

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Ten alleles (five M and five N alleles) of the MN blood group system with normal antigenicity were found by sequencing the glycophorin A (GPA) gene. This study demonstrates the systematic classification of these alleles to major or minor variations of the standard alleles. GPA-specific fragments ranging from 150 to 3.8 kb in length were amplified from the templates, and exons 1-7 and introns 1-6 were sequenced. The data were analyzed phylogenetically to classify these alleles into major groups or clusters. The ten alleles were grouped into four major clusters M10X (M101-M103), M20X (M201 and M202), N10X (N101-N104) and N20X (N201), where 'X' represents a digit indicating minor variations. This grouping was supported by phylogenetic analysis. The cluster system of GPA alleles is highly informative for genetic screening.

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“…The MN blood group system can be genotyped serologically or by allele-specific PCR [8, 9, 10], reverse dot blot hybridization [11], PCR-restriction fragment length polymorphism (RFLP), or single-strand conformational polymorphism (SSCP) techniques [12]. Previously, detecting M and N alleles by PCR-RFLP and PCR-SSCP methods, we found a nucleotide (G/T) substitution in intron 1 of the M alleles, and classified conventional M alleles into two alleles, provisionally named M G and M T [12].…”

Section: Introductionmentioning

confidence: 99%

Classification of Standard Alleles of the MN Blood Group System

2000

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Background and Objectives: We had classified the M alleles of the MN blood group system into two subtypes, M^G (standard M) and M^T, based on a G/T substitution in intron 1 of the glycophorin A (GPA) gene. This study provides further study on nucleotide sequences of M^G, M^T and N alleles to profile the new allele, M^T. Materials and Methods: M^G, M^T and N alleles of the GPA gene were amplified using GPA gene-specific primers to avoid co-amplification of the genes of glycophorins B and E. Then the 5′-flanking region, exons 1–7 and introns 1–4 of the alleles were analyzed by DNA sequencing. Results: There were 17 nucleotide substitutions and deletions between M^G (standard M) and N alleles. Ten of the M^T nucleotides were M^G-type but the other 7 were N-type. M^T allele also showed one base change and one deletion that were observed in neither the M^G nor the N allele. Moreover, we found nucleotide substitutions within each allele, allowing further classification of the alleles. Conclusion: By the sequence data of M^G, M^T and N alleles, the three alleles could be further classified into M101 and M102, M201 and M202, and N101 and N102, respectively.

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An improved method for MN genotyping by the polymerase chain reaction

Cited by 11 publications

References 4 publications

PCR-based genotyping of MNSs blood group: Subtyping of M allele to MG and MT

PCR-based genotyping of MNSs blood group: Subtyping of M allele to MG and MT

Systematic classification of alleles of the glycophorin A (MN blood group) gene

Classification of Standard Alleles of the MN Blood Group System

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