Extension of base mispairs byTaqDNA polymerase: implications for single nucleotide discrimination in PCR
نویسندگان
چکیده
منابع مشابه
Extension of base mispairs by Taq DNA polymerase: implications for single nucleotide discrimination in PCR.
Thermus aquaticus (Taq) DNA polymerase was used to measure the extension efficiency for all configurations of matched and mismatched base pairs at template-primer 3'-termini. The transition mispairs, A(primer).C, C.A, G.T, and T.G were extended 10(-3) to 10(-4)-fold less efficiently than their correctly paired counterparts. Relative efficiencies for extending transversion mispairs were 10(-4) t...
متن کاملA Microfluidic Device for Detection of Single Nucleotide Polymorphisms by Allele Specific Single Base Extension
We present a microfluidic device with integrated temperature sensors and heaters for single nucleotide polymorphism (SNP) detection. The allele specific primer is immediately annealed next to the polymorphic site, and extended by a single base. The extension product is then purified by solid phase capture, and subsequently released via chemical cleavage. After desalting, the released product is...
متن کاملA microsphere-based assay for multiplexed single nucleotide polymorphism analysis using single base chain extension.
A rapid, high throughput readout for single-nucleotide polymorphism (SNP) analysis was developed employing single base chain extension and cytometric analysis of an array of fluorescent microspheres. An array of fluorescent microspheres was coupled with uniquely identifying sequences, termed complementary ZipCodes (cZipCodes), which allowed for multiplexing possibilities. For a given assay, que...
متن کاملDiscrimination against purine-pyrimidine mispairs in the polymerase active site of DNA polymerase I: a structural explanation.
We previously identified five derivatives of Klenow fragment DNA polymerase that have lower fidelity because of amino acid substitutions in the polymerase active site. One of these has alanine substituted for the invariant Glu-710, whose side chain interacts with the deoxyribose of the incoming dNTP. Here we show that the E710A enzyme has reduced fidelity for five of the 12 possible mismatches....
متن کاملAllelic Discrimination for Single Nucleotide Polymor-
The scavenger receptor class B type 1 (SR-BI), a multiligand receptor, appears to be a physiologically relevant HDL receptor in rodents (1, 2). To determine its role in humans, the human SRB1 gene has been characterized (3, 4) and its genetic variation investigated in a Caucasian population (4 ). We have reported three variants, at exons 1 and 8 and intron 5, with allele frequencies .0.1 that h...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: Nucleic Acids Research
سال: 1992
ISSN: 0305-1048,1362-4962
DOI: 10.1093/nar/20.17.4567