In this proof-of-principle study we show that linear amplification is possible over a wide range of amplification cycles. Professor Larry Wangh, Ph.D., and his laboratory colleagues have invented a unique DNA amplification and analysis technique called “Linear After The Exponential (LATE) PCR (Polymerase Chain Reaction).” LATE-PCR is an advanced form of asymmetric PCR which begins by generating double-stranded DNA molecules but then switches to efficient amplification of single-stranded molecules. Aims To verify monoplex and multiplex gene‐specific linear‐after‐the‐exponential polymerase chain reaction (LATE‐PCR) assays for identifying 17 microbial pathogens (i.e., Klebsiella sp., Acinetobacter baumannii, Staphylococcus aureus, Enterobacter sp., Pseudomonas aeruginosa, coagulase negative staphylococci, Enterococcus sp., Candida sp.) This method, however, is difficult to optimize, often inefficient, and tends to promote nonspecific amplification. 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. During the early cycles of PCR, the amplification is exponential. When tested against synthetic targets, the assay … Due to the slow (arithmetic) amplification later in the reaction (after the limiting primer has been used up) extra cycles of PCR are required. Our preferred strategy, Linear-After-The-Exponential (LATE)-PCR, uses primers that are deliberately designed for use at unequal concentrations, such that the concentration-adjusted Tm of the limiting primer (TmL) is at least as high as the concentration-adjusted melting temperature of the excess primer (TmX) (i.e., TmL - TmX 2 O) (4, 8). Because of the properties of LATE-PCR, each reaction produces large amount of specific, single-stranded DNA, which can then be probed with a sequence-specific probe. Here we provide details of linear-after-the-exponential-PCR (LATE-PCR), a method similar to asymmetric PCR in the use of primers at different concentrations, but with novel design criteria to ensure high efficiency and specificity. 3. LATE-PCR also introduces new probe design criteria that uncouple hybridization probe detection from primer … Linear-after-the-exponential (LATE)-PCR describes a novel approach to asymmetric PCR which uses adjusted melting temperatures of the limited primer to increase PCR efficiency. What is the physical origin of the saturation and why is the explanation important for PCR design? 4. 1 ways to abbreviate Linear-after-the-exponential-PCR updated 2020. Kenneth E. Pierce, J. Aquiles Sanchez, John E. Rice, and Lawrence J. Wangh PNAS, 2005, 102 (24): 8609–8614 . How to abbreviate Linear-after-the-exponential-PCR? Gerald Thulbourn 2017-12-05T09:42:49+00:00 December 5th, 2017 | A form of asymmetric PCR that uses primer pairs deliberately designed for use at unequal concentrations (Pierce, 2003; Sanchez, 2004). PCR assays, regardless of primer ratio. signatures, linear-after-the-exponential-PCR, septicaemia, sequence-specific low temperature probes. Our preferred strategy, Linear-After-The-Exponential (LATE)-PCR, uses primers that are deliberately designed for use at unequal concentrations, such that the concentration-adjusted T m of the limiting primer (T m L) is at least as high as the concentration-adjusted melting temperature of the excess primer (T m X) (i.e., T m L - T m X ≥ 0) (4, 8). Linear-after-the-exponential (LATE)–PCR is a novel, highly robust, asymmetric PCR method that uses unequal concentrations of primers to generate large amounts of ssDNA together with a small fixed amount of double-stranded DNA (dsDNA) [29,30]. Aims: A novel molecular assay for the detection of foot-and-mouth disease virus (FMDV) was developed using linear-after-the-exponential polymerase chain reaction (LATE-PCR). recent modification on this process, known as Linear-After-The-Exponential-PCR (LATE-PCR), uses a limiting primer with a higher melting temperature (Tm) than the excess primer to maintain reaction efficiency as the limiting primer concentration decreases mid-reaction. - "Linear-after-the-exponential polymerase chain reaction and allied technologies. LATE (Linear After The Exponential)-PCR. Linear-After-The-Exponential (LATE)-PCR and Its Allied Technologies for highly informative, highly accurate DNA amplification and application of these technologies for design and construction of diagnostic assays in many fields. Real-time detection strategies for rapid, reliable diagnosis from single cells." As amplicons from LATE-PCR contain a large amount of ssDNA, pyrosequencing can be performed on the amplicons … A successful LATE-PCR was achieved by using a common Taq DNA polymerase in high pH buffer (HpH-buffer). To simplify the process of a pyrosequencing protocol, we proposed an improved linear-after-the-exponential (LATE)-PCR by employing whole blood as the starting material. LATE - Linear-After-The-Exponential. During the later stages of PCR, saturation behavior is observed, and the amplification efficiency of PCR decreases with each successive cycle. Methods and results: Pilot experiments using synthetic DNA targets demonstrated the ability of LATE-PCR to quantify initial target concentration through endpoint detection. The boxes labeled 1, 2, and 3 indicate diagnostic limits for those genotypes that were established by previously tested samples. LATE-PCR - Linear-After-the-Exponential Polymerase Chain Reaction. Here linear-after-the-exponential (LATE)-PCR is introduced as an efficient and potentially automatable method of directly amplifying single-stranded DNA for pyrosequencing, thereby eliminating the need for solid-phase sample preparation and reducing the risk of laboratory contamination. 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. The most popular abbreviation for Linear-after-the-exponential-PCR is: LATE-PCR The resulting amplification strategy, termed Linear-After-The-Exponential PCR (LATE-PCR) is efficient and specific (5,6). Linear-after-the-exponential (LATE)-PCR describes a novel approach to asymmetric PCR which uses adjusted melting temperatures of the limited primer to increase PCR efficiency. Traditional asymmetric PCR uses conventional PCR primers at unequal concentrations to generate single-stranded DNA. Asymmetric PCR differs from regular PCR by the excessive amount of primers for a chosen strand. Linear-After-the-Exponential Polymerase Chain Reaction - How is Linear-After-the-Exponential Polymerase Chain Reaction abbreviated? Aims: A novel molecular assay for the detection of foot-and-mouth disease virus (FMDV) was developed using linear-after-the-exponential polymerase chain reaction (LATE-PCR). GPS Global Positioning System; ACCA Automated Cloud Cover Assessment; CFA Cyclopropane fatty acids; BSCL Berardinelli-Seip congenital lipodystrophy; AP Alloy Primer; AP Adhesion Promoter; AGS Aicardi-Goutieres syndrome; ACP Annealing Control Primer; ACD Active case detection; AABB Axis-Aligned Bounding Boxes; ARMP Agricultural Research Management … Aims: A novel molecular assay for the detection of foot‐and‐mouth disease virus (FMDV) was developed using linear‐after‐the‐exponential polymerase chain reaction (LATE‐PCR). Fig. Looking for abbreviations of LATE-PCR? These results show that detection during PCR at 50C fluorescence will provide close to maximum fluorescence with that target (arrow), but only background fluorescence with the mismatched target. It is Linear-After-the-Exponential Polymerase Chain Reaction. Unlike typical asymmetric PCR, LATE-PCR, amplification is efficient due to improved primer design (Pierce, 2005). LATE-PCR generates single-stranded products with predictable kinetics for many cycles be-yond the exponential phase. Aims: To verify monoplex and multiplex gene-specific linear-after-the-exponential polymerase chain reaction (LATE-PCR) assays for identifying 17 microbial pathogens (i.e., Klebsiella sp., Acinetobacter baumannii, Staphylococcus aureus, Enterobacter sp., Pseudomonas aeruginosa, coagulase negative staphylococci, Enterococcus sp., Candida sp.) This paper describes the design and verification of a novel assay for detection of African swine fever virus (ASFV) based on Linear-After-The-Exponential PCR (LATE-PCR). Linear-After-The-Exponential (LATE)-PCR describes a new paradigm for primer design that renders assays as efficient as sym. 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. An alternative approach, Linear-After-The-Exponential (LATE)-PCR, solves these problems by using primer pairs deliberately designed for use at unequal concentrations. Linear-After-the-Exponential Polymerase Chain Reaction listed as LATE-PCR . Linear-After-The-Exponential PCR (LATE-PCR) Talk at the 2nd Nucleic Acid Quantification Meeting in London 2003: by Larry Wangh Paper: Primer design criteria for high yields of specific single-stranded DNA and improved real-time detection. Fig. Methods and Results: Pilot experiments using synthetic DNA targets demonstrated the ability of LATE‐PCR to quantify initial target concentration through endpoint detection. LATE-PCR generates single-stranded products with predictable kinetics for many cycles beyond the exponential phase. Methods and Results: Pilot experiments using synthetic DNA targets demon- Viele übersetzte Beispielsätze mit "linear-after-the-exponential" – Deutsch-Englisch Wörterbuch und Suchmaschine für Millionen von Deutsch-Übersetzungen. 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