Generation of competitor DNA fragments for quantitative PCR.
نویسندگان
چکیده
منابع مشابه
PCR amplification of long DNA fragments.
The polymerase chain reaction (PCR) has recently evolved as a standard laboratory technique, popular in all areas of molecular biology research. However, the technique still has two limitations: the relatively low fidelity of Taq polymerase when compared with other polymerases (1), and its inability to efficiently amplify fragments higher than 3 kbp (2, 3). Although these two issues are irrelev...
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Accurate and rapid diagnosis of human cytomegalovirus (HCMV) disease in immunocompromised patients has remained as a challenge. Quantitative competitive PCR (QC-PCR) methods for detection of HCMV in these individuals have improved the positive and negative predictive values of PCR for diagnosis of HCMV disease. In this study we used QC-PCR assay, using a co-amplified DNA standard, to quantitate...
متن کاملCompetitive RT-PCR: Preparation of Competitor RNA.
INTRODUCTION The first step in competitive RT-PCR is the synthesis and purification of the synthetic competitor. This is an RNA molecule designed to be reverse-transcribed and PCR-amplified with the same efficiency as the endogenous transcript of interest. Once the competitor molecule has been prepared, as described in this protocol, competitive PCR can be carried out, as described in Comptetit...
متن کاملChallenges to Design and Develop of DNA Aptamers for Protein Targets. I. Optimization of Asymmetric PCR for Generation of a Single Stranded DNA Library
Aptamers, or single stranded oligonucleotides, are produced by systematic evolution of ligands by exponential enrichment, abbreviated as SELEX. In the amplification and regeneration step of SELEX technique, dsDNA is conversed to ssDNA. Asymmetric PCR is one of the methods used for the generation of ssDNA. The purpose of this study was to design a random DNA library for selection of aptamers wit...
متن کاملChallenges to Design and Develop of DNA Aptamers for Protein Targets. I. Optimization of Asymmetric PCR for Generation of a Single Stranded DNA Library
Aptamers, or single stranded oligonucleotides, are produced by systematic evolution of ligands by exponential enrichment, abbreviated as SELEX. In the amplification and regeneration step of SELEX technique, dsDNA is conversed to ssDNA. Asymmetric PCR is one of the methods used for the generation of ssDNA. The purpose of this study was to design a random DNA library for selection of aptamers wit...
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ژورنال
عنوان ژورنال: Genome Research
سال: 1991
ISSN: 1088-9051
DOI: 10.1101/gr.1.2.136