Cloning-Free PCR-Based Allele Replacement Methods
نویسندگان
چکیده
منابع مشابه
Cloning-free PCR-based allele replacement methods.
Efficient homologous recombination permits the directed introduction of specific mutations into the yeast genome. Here we describe a cloning-free, PCR-based allele replacement method that simplifies allele transfer between yeast strains. The desired allele from one strain is amplified by PCR, along with a selectable/counterselectable marker. After transformation, the resident allele in the targ...
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In many DNA amplification reactions, extra fragments arising from nonspecific mispriming, a mixture of target templates or allelic variation may be generated in addition to the desired product. Some nonspecific bands can be reduced by more stringent annealing conditions, but in cases of allelic variation or other mixed targets, the amplification products may be of the same length but differ by ...
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Cloning of polymerase chain reaction (PCR) products into plasmid vectors by conventional methods, relying either on recognition sequences built into the primers or taking advantage of the template-independent terminal transferase activity of Taq DNA polymerase to add an extra adenine to the 3′ end of the product, are often difficult (1,10,12). In recent years, numerous cloning strategies (10,13...
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Determination of allele frequency in pooled DNA samples is a powerful and efficient tool for large-scale association studies. In this study, we tested and compared three PCR-based methods for accuracy, reproducibility, cost, and convenience. The methods compared were: (i) real-time PCR with allele-specific primers, (ii) real-time PCR with allele-specific TaqMan probes, and (iii) quantitative se...
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BACKGROUND In high-throughput demanding fields, such as biotechnology and structural biology, molecular cloning is an essential tool in obtaining high yields of recombinant protein. Here, we address recently developed restriction-free methods in cloning, and present a more cost-efficient protocol that has been optimized to improve both cloning and clone screening. RESULTS In our case study, t...
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ژورنال
عنوان ژورنال: Genome Research
سال: 1997
ISSN: 1088-9051,1549-5469
DOI: 10.1101/gr.7.12.1174