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Dergiler / Turkish Journal of Hematology / 2001 / Cilt: 18 - Sayı: 2

Rapid identification of family-specific mutations in the factor VIII gene by one-step DGGE

Anıl Ayşe TİMUR
Davut ALBAYRAK
S. Hande ÇAĞLAYAN
Sayfa
89–93
DOI
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One-step DGGE is a promising rapid method for the carrier detection and prenatal diagnosis in haemophilia A families when immediate results are required and when polymorphic markers fail to give information.

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Amaç & Hipotez:A one-step denaturing gradient gel electrophoresis (DGGE) strategy for the rapid detection of mutations in the factor VIII gene of haemophilia A patients is described.
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Temel Bulgular & Veri:All coding (except the middle part of exon 14) and flanking intronic regions of the gene corresponding to approximately 6.6 kb were amplified in 27 fragments using four PCR programs.
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Sonuç & Katkı:One-step DGGE is a promising rapid method for the carrier detection and prenatal diagnosis in haemophilia A families when immediate results are required and when polymorphic markers fail to give information.
🔬Tür:Araştırma Makalesi
📍Kapsam:Genişletilmiş Akademik Örneklem
⚙️Değişkenler:Hemofili A · Elektroforez · jel

Özet

A one-step denaturing gradient gel electrophoresis (DGGE) strategy for the rapid detection of mutations in the factor VIII gene of haemophilia A patients is described. All coding (except the middle part of exon 14) and flanking intronic regions of the gene corresponding to approximately 6.6 kb were amplified in 27 fragments using four PCR programs. Heteroduplex formation was performed for each fragment. A common denaturant gradient gel (35-65%) was chosen that allowed the simultaneous analysis of all PCR amplified regions on a single gel and run for 3.5 h at 160 V. This method was implemented for a patient whose family was seeking carrier determinations. An abnormal pattern was detected in exon 23 and the family-specific mutation was found by subsequent DNA sequencing. One-step DGGE is a promising rapid method for the carrier detection and prenatal diagnosis in haemophilia A families when immediate results are required and when polymorphic markers fail to give information.

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Bu Makaleye Atıf Yap

TİMUR, A. A., ALBAYRAK, D., ÇAĞLAYAN, S. H. (2001). Rapid identification of family-specific mutations in the factor VIII gene by one-step DGGE. Turkish Journal of Hematology, Cilt: 18 - Sayı: 2, 89–93. https://ajindex.com/articles/412976

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Kaynakça (12)

  1. Kemball-Cook G, Tuddenham, EGD, Wacey AI. Thefactor VIII structure and mutation resource site:HAMSTeRS version 4. Nucleic Acids Res 1998
  2. Lakich D, Kazazian, HHJR, Antonarakis SE, Gitsc-hier J. Inversions disrupting the factor VIII gene area common cause of severe heamophilia A. Nat Ge-net 1993
  3. Cotton RG, Rodrigues NR, Campbell RD. Reactivityof cytosine and thymine in single-base pair mismatc-hes with hydroxylamine and osmium tetraoxide andits applications to the study of mutations. Proc NatlAcad Sci USA 1988
  4. Freson K, Peerlinck K, Aguirre T, Arnout, J, Vermy-len, J, Cassiman J-J, Matthijs G. Fluorescent chemi-cal cleavage of mismatches for efficient screening ofthe factor VIII gene. Hum Mutat 1998
  5. Orita M, Suzuki Y, Sekiya T, Hayashi K. Rapid andsensitive detection of mutations and DNA polymorp-hisms using the polymerase chain reaction. Geno-mics 1989
  6. Pieneman WC, Deutz-Terlouw PP, Reitsma PH, Bri-et E. Screening for mutations in haemophilia A pati-ents by multiplex PCR-SSCP, Southern blotting andRNA analysis: The detection of a genetic abnorma-lity in the factor VIII gene in 30 out of 35 patients.Br J Haematol 1995
  7. Myers RM, Maniatis T, Lerman LS. Detection andlocalization of single base changes by denaturing gra-dient gel electrophoresis. Methods Enzymol 1987
  8. Higuchi M, Antonarakis SE, Kasch L, Oldenburg J,Economou-Petersen E, Olek K, Arai M, Inaba H,Kazazian HHJR. Molecular characterization of mild-to-moderate hemophilia A: Detection of the mutati-on in 25 of 29 patients by denaturing gradient gelelectrophoresis. Proc Natl Acad Sci USA1991
  9. Schwaab R, Oldenburg J, Schwaab U, JohnsonDJD, Schmidt W, Olek K. Characterization of muta-tions within the factor VIII gene of 73 unrelated mildand moderate haemophiliacs. Br J Haematol1995
  10. Sheffield VC, Cox DR, Lerman LS, Myers RM. At-tachment of a 40-base-pair G+C-rich sequence (GC-clamp) to genomic DNA fragments by the polymera-se chain reaction results in improved detection ofsingle base changes. Proc Natl Acad Sci USA 1989
  11. Schwaab R, Oldenburg J, Lalloz MRA, Schwaab U,Pemberton S, Hanfland P, Brackmann HH, Tudden-ham EGD, Michaelides K. Factor VIII gene mutati-ons found by a comparative study of SSCP, DGGEand CMC and their analysis on a molecular model offactor VIII protein. Hum Genet 1997
  12. Timur AA, Gürgey A, Aktuğlu G, Kavaklı K, Cana-tan D, Olek K, Çağlayan SH. Mo