Thermal profile with alternately raised and lowered annealing temperature improves the PCR amplification using highly degenerate primers.
نویسندگان
چکیده
منابع مشابه
Amplification of Hsp90 homologs from plant-parasitic nematodes using degenerate primers and ramped annealing PCR.
Procedures for using PCR to amplify novel members of gene families from genomic DNA frequently involve the use of degenerate primers. Of key importance is the ability to produce ample quantities of specific PCR product while minimizing or eliminating nonspecific side reactions. Primer design protocols and software are improving, but finding the optimal amplification conditions often involves a ...
متن کاملMAD-DPD: designing highly degenerate primers with maximum amplification specificity.
This work introduces minimum accumulative degeneracy, a variant of the degenerate primer design problem, which is particularly useful when a large number of sequences are to be covered by a set of restricted number of primers. A primer set, which is designed on a minimum accumulative degeneracy basis, especially helps to reduce nonspecific PCR amplification of undesired DNA fragments, as fewer ...
متن کاملSelecting Degenerate Multiplex PCR Primers
Single Nucleotide Polymorphism (SNP) Genotyping is an important molecular genetics technique in the early stages of producing results that will be useful in the medical field. One of the proposed methods for performing SNP Genotyping requires amplifying regions of DNA surrounding a large number of SNP loci. In order to automate a portion of this method and make the use of SNP Genotyping more wi...
متن کاملPlasmid DNA sequencing using highly degenerate oligonucleotides as primers.
Sequencing double-stranded DNA templates has become an efficient procedure (1) being much less time-consuming than the preparation and subsequent sequencing of inserts in phage singlestranded DNA. The ability to use highly degenerate oligonucleotides instead of sequence-specific primers would make this method even more powerful for the rapid sequencing ofDNA that codes for a protein for which a...
متن کامل5'-degenerate 3'-dideoxy-terminated competitors of PCR primers increase specificity of amplification.
Amplification of a product in PCR with specific primers may be viewed as an artificial Darwinian-type "selection of the fittest". In other selective systems, such as general evolution, immune system and probably brain cortex, the stringency of selection is not absolute but rather degenerate, with selection of many highly fit units, not limited, however, to only the fittest. In PCR also, anneali...
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: Acta Biochimica Polonica
سال: 1998
ISSN: 1734-154X,0001-527X
DOI: 10.18388/abp.1998_4262