The major limitation of this technique is the requirement for large amounts of input DNA, consequently precluding the analysis of paraffin-embedded tissues. The PCR method is useful in situations in which the amount of DNA sample is limited, such as in forensics and prenatal testing, or in which the quality of the DNA sample is poor. Southern blot analysis can be used to detect point mutations that disrupt restriction sites, as well as larger structural changes, such as insertions, deletions, and translocations. This technique detects specific DNA sequences and therefore is also. Initially, an electrophoretic procedure is used to separate molecules (or protein and nucleic acid fragments). In other areas, PCR is being used to detect and characterize microbial pathogens and to characterize mutations associated with carcinogenesis. Edwin Southern developed one of the first blotting techniques called the Southern blot. All blotting techniques share a similar workflow. In addition, the PCR technique can be applied to polymorphism analysis to provide diagnosis by linkage analysis. Less often, gene sequencing of a PCR product is used to rapidly identify a mutation. DNA samples can be obtained from tissue or. The Southern blot method may also be used to determine the molecular weight of restriction fragments and to measure relative amounts of DNA in different samples. After PCR, mutations producing single-gene disorders can be detected by several different methods, including endonuclease digestion and gel electrophoresis (applicable when a mutation affects an endonuclease recognition site), gel electrophoresis (used for detection of deletions), and hybridization to an oligonucleotide probe specific for a mutation. Southern blotting is a molecular biology technique used for the detection of a specific DNA sequence in large, complex samples of DNA. The technique is being used for rapid prenatal diagnosis and carrier testing of several inherited disorders. Southern blotting is used in molecular biology for the identification of proteins and nucleic acids and is widely used for diagnostic purposes. The polymerase chain reaction (PCR) is a rapid method for generating a 10(6)- to 10(7)-fold increase in the number of copies of a discrete DNA or RNA sequence.
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