Linear-after-the-exponential (LATE)-PCR: Improved asymmetric PCR for quantitative DNA-analysis
نویسندگان
چکیده
منابع مشابه
Linear-after-the-exponential (LATE)-PCR: an advanced method of asymmetric PCR and its uses in quantitative real-time analysis.
Conventional asymmetric PCR is inefficient and difficult to optimize because limiting the concentration of one primer lowers its melting temperature below the reaction annealing temperature. Linear-After-The-Exponential (LATE)-PCR describes a new paradigm for primer design that renders assays as efficient as symmetric PCR assays, regardless of primer ratio. LATE-PCR generates single-stranded pr...
متن کاملDirect amplification of single-stranded DNA for pyrosequencing using linear-after-the-exponential (LATE)-PCR.
Pyrosequencing is a highly effective method for quantitatively genotyping short genetic sequences, but it currently is hampered by a labor-intensive sample preparation process designed to isolate single-stranded DNA from double-stranded products generated by conventional PCR. Here linear-after-the-exponential (LATE)-PCR is introduced as an efficient and potentially automatable method of directl...
متن کاملLinear-After-The-Exponential (LATE)-PCR: primer design criteria for high yields of specific single-stranded DNA and improved real-time detection.
Traditional asymmetric PCR uses conventional PCR primers at unequal concentrations to generate single-stranded DNA. This method, however, is difficult to optimize, often inefficient, and tends to promote nonspecific amplification. An alternative approach, Linear-After-The-Exponential (LATE)-PCR, solves these problems by using primer pairs deliberately designed for use at unequal concentrations....
متن کاملImproved resolution of asymmetric-PCR SSCP products.
amide gel electrophoresis (PAGE) on a 180× 160× 1.5-mm 10% polyacrylamide gel. The separated DNA strands were subsequently visualized by staining with either EtdBr or silver (2). Separation of the DNA strands before electrophoresis using streptavidin paramagnetic beads consistently yielded single-stranded products upon SSCP analysis (Figure 1). Treatment of DNA strands with S1 nuclease before S...
متن کاملDesign and verification of a highly reliable Linear-After-The-Exponential PCR (LATE-PCR) assay for the detection of African swine fever virus.
African swine fever virus (ASFV) is a highly pathogenic DNA virus that is the causative agent of African swine fever (ASF), an infectious disease of domestic and wild pigs of all breeds and ages, causing a range of syndromes. Acute disease is characterized by high fever, haemorrhages in the reticuloendothelial system, and a high mortality rate. A powerful novel diagnostic assay based on the Lin...
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ژورنال
عنوان ژورنال: Forensic Science International: Genetics Supplement Series
سال: 2015
ISSN: 1875-1768
DOI: 10.1016/j.fsigss.2015.09.246