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Biochemistry General 699ace40

Which test uses oligomer with single base pair substitution:

A
PCR
B
RFLP
C
Error coded mutation analysis
D
None of the above
High-Yield Explanation
RFLP [Ref: J Polymorphisms are inherited differences found among the individuals in a population. Restriction Fragment Length Polymorphism (RFLP) is a difference in homologous DNA sequences that can be detected by the presence of fragments of different lengths after digestion of the DNA samples in question with specific restriction endonucleases. RFLP, as a molecular marker, is specific to a single clone/restriction enzyme combination. An RFLP probe is a labeled DNA sequence that hybridizes with one or more fragments of the digested DNA sample after they were separated by gel electrophoresis, thus revealing a unique blotting pattern characteristic to a specific genotype at a specific locus. Sho, single- or low-copy genomic DNA or cDNA clones are typically used as RFLP probes. RFLPs are visualized by digesting DNA from different individuals with a restriction enzyme, followed by gel electrophoresis to separate fragments according to size, then blotting and hybridization to a labeled probe that identifies the locus under investigation. An RFLP is demonstrated whenever the Southern blot pattern obtained with one individual is different from the one obtained with another individual. Harrison 17/e p390 writes-"Southern blotting is used to analyze whether genes have been deleted or rearranged. It is also used to detect restriction fragment length polymorphisms (RFLPs). Genornic DNA is digested with restriction endonucleases and separated by gel electrophoresis. Individual .fragments can then be transferred to a membrane and detected after hybridization with specific radioactive DNA probes. Because single base pair mismatches can disrupt the hybridization of sho DNA probes (oligonucleotides), a variation of the Southern blot, termed oligonucleotide-specific hybridization (OSH), uses sho oligonucleotides to distinguish normal from mutant genes." RFLP is an impoant tool in genome mapping, localization of genes for genetic disorders, determination of risk for disease, and paternity testing

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