Quantitative multiplexing analysis of PCR-amplified ribosomal RNA genes by hierarchical oligonucleotide primer extension reaction
نویسندگان
چکیده
منابع مشابه
Quantitative multiplexing analysis of PCR-amplified ribosomal RNA genes by hierarchical oligonucleotide primer extension reaction
A method, termed hierarchical oligonucleotide primer extension (HOPE), is developed for quantitative, multiplexing detection of DNA targets present in PCR-amplified community 16S rRNA genes. It involves strand extension reaction and multiple oligonucleotide primers modified with different lengths of polyA at the 5' end and targeting 16S rRNA genes at different phylogenetic specificities. On ann...
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1. Phosphorylate oligonucleotide probe: a. Oligonucleotide primer (5 – 7 pmoles or 60 ng) 1 μL b. dH2O 6.5 μL c. 10X kinase buffer 1.5 μL d. T4 PNK (~10 U) 1 μL e. [γ-P]ATP (7000 Ci/mmole) 2 μL 2. Stop the kinase reaction by adding 500 μL of TE (pH 7.6) 3. Add 25 μg of carrier RNA. 4. Phenol/Chloroform extract 2X 5. Ethanol precipitate with sodium acetate (pH 5.2) 6. Wash with 70% ethanol and r...
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We have developed a new primer design strategy for PCR amplification of distantly related gene sequences based on consensus-degenerate hybrid oligonucleotide primers (CODEHOPs). An interactive program has been written to design CODEHOP PCR primers from conserved blocks of amino acids within multiply-aligned protein sequences. Each CODEHOP consists of a pool of related primers containing all pos...
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19 § These authors contributed equally to this work 20 21 22 2 ABSTRACT 1 2 One of the first steps in characterizing an ecosystem is to describe the organisms 3 inhabiting it. For microbial studies, experimental limitations have hindered the ability to depict 4 these diverse communities. Here we describe oligonucleotide fingerprinting of ribosomal RNA 5 genes (OFRG), a method that permits the i...
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ژورنال
عنوان ژورنال: Nucleic Acids Research
سال: 2007
ISSN: 1362-4962,0305-1048
DOI: 10.1093/nar/gkm413