Detection of point mutations by non-isotopic single strand conformation polymorphism
نویسندگان
چکیده
منابع مشابه
Detection of point mutations by non-isotopic single strand conformation polymorphism.
We have developed a procedure for nonradioactive single strand conformation polymorphism analysis and applied it to the detection of point mutations in the human tumor suppressor gene p53. The protocol does not require any particular facilities or equipment, such as radioactive handling, large gel units for sequencing, or a semiautomated electrophoresis system. This technique consists of amplif...
متن کاملMutation Detection by Single Strand Conformation Polymorphism and Heteroduplex Analysis
Single strand conformation polymorphism (SSCP) and heteroduplex analysis (HDA) are two of the most popular electrophoresis-based mutation detection methods. Coupled to DNA amplification of the sequence to be analyzed, these techniques have become the methods of choice for a number of molecular diagnostic laboratories. This can be explained mainly by the numerous advantages, namely their technic...
متن کاملDetection of Mutations by Single-strand Conformational Polymorphism.
MATERIALS 10x Amplification buffer Include 0.01% (w/v) gelatin in the buffer. 10x TBE electrophoresis buffer Use TBE at a working strength of 1x (89 mM Tris-borate, 2 mM EDTA) for polyacrylamide gel electrophoresis. 6x Gel-loading buffer I Formamide loading buffer Human genomic DNA to be screened for point mutations Dissolve the DNA at 10 μg/ml in TE (pH 7.6). Oligonucleotide primers, forward a...
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Medium chain acyl-CoA dehydrogenase (MCAD) deficiency is a common inherited metabolic disorder affecting fatty acid beta oxidation. Identification of carriers is important since the disease can be fatal and is readily treatable once diagnosed. Twelve molecular defects have been identified in the MCAD gene; however, a single highly prevalent mutation, A985G, accounts for > 90% of mutant alleles ...
متن کاملnon-radioactive single-strand conformation polymorphism (sscp) analysis of relatively long pcr products
the sscp technique is based on the appearance of new “refolding” conformations during electro-phoresis due to mutation. in order to develop a simple, non-radioactive sscp analysis method so that it can reliably detect single nucleotide changes in pcr products up to 500 bp in length, extensive optimisation trials were performed . the best separation of sscp bands of pcr products up to 500 bp in ...
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ژورنال
عنوان ژورنال: Brazilian Journal of Medical and Biological Research
سال: 1999
ISSN: 0100-879X
DOI: 10.1590/s0100-879x1999000100008