Efficient amplification using ‘megaprimer’ by asymmetric polymerase chain reaction
نویسندگان
چکیده
منابع مشابه
SPECIFIC AMPLIFICATION OF ASPERGILLUS FUMlGATUS DNA BY POLYMERASE CHAIN REACTION
Invasive aspergillosis (1 is a life-threatening condition in immunocompromised patients. An early diagnosis is of great importance because early treatment may resolve this potentially fatal infection. Recently, the polymerase chain reaction (PCR) has been used successfully in detecting specific DNA of several pathogen. In this study, nested PCR was used to detect DNA specific for A!.pergiflus s...
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The "megaprimer" method (1) based on polymerase chain reaction (PCR) is one of the simplest and most versatile procedures of site-specific in vitro mutagenesis available to date. The method utilizes three oligonucleotide primers and two rounds of PCR performed on a DNA template containing the cloned gene that is to be mutated. The rationale of the method is shown schematically in Fig. 1 where A...
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Site-directed mutagenesis has become an essential tool in the investigation of gene structure and function as well as the products of gene expression, namely RNA and protein. Selected positions in a DNA sequence are changed, and the eVects of the mutations are studied in vitro and/or in vivo. Although many diVerent mutagenic strategies are available [1], the simplicity, cost eVectiveness, and a...
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invasive aspergillosis (1 is a life-threatening condition in immunocompromised patients. an early diagnosis is of great importance because early treatment may resolve this potentially fatal infection. recently, the polymerase chain reaction (pcr) has been used successfully in detecting specific dna of several pathogen. in this study, nested pcr was used to detect dna specific for a!.pergiflus s...
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The polymerase chain reaction (PCR) and Q/3 replicase are two methods in which nucleic acid polymerases are used for amplification. Although these approaches share many similar problems concerning target contamination and probe specificity, they differ dramatically in their mechanisms of action and modes of application.The PCR method amplifies target sequences between two priming oligonucleotid...
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ژورنال
عنوان ژورنال: Nucleic Acids Research
سال: 1995
ISSN: 0305-1048,1362-4962
DOI: 10.1093/nar/23.21.4530