Rapid Screening of Mutant N-ras Alleles by Analysis of PCR-Induced Restriction Sites: Allele Specific Restriction Analysis (ASRA).
作者： Alison V. Todd , Harry J. Iland
DOI: 10.3109/10428199109107917
关键词: DNA 、 Mutant 、 Molecular biology 、 Gene 、 Gene mutation 、 Biology 、 Restriction site 、 Wild type 、 Genetics 、 Primer (molecular biology) 、 Restriction fragment length polymorphism
摘要: We have developed a rapid screening method for analysis of codon 12, 13 and 61 N-ras gene mutations, since these mutations been observed in approximately 25% patients with acute myeloid leukemia myelodysplastic syndromes. The method, termed allele specific restriction (ASRA), involves polymerase chain reaction amplification DNA or RNA using mismatched primer which introduces appropriately positioned base substitutions creates site provided the adjacent sequence is normal. Simultaneous codons 12 also possible by use multiprimer mixture. Resistance amplified product to digestion indicates presence mutation original template. Since ASRA allows simultaneous mutant wild type sequences RNA, an estimate ratio copies relative expression alleles can be obtained heterozygous individuals.
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