DGGE/TGGE (Denaturing Gradient Gel Electrophoresis/Temperature Gradient Gel Electrophoresis) analysis method of specific 18S rDNA (Deoxyribose Nucleic Acid) fragment without being based on GC clamp strategy

Abstract

The invention provides a DGGE / TGGE (Denaturing Gradient Gel Electrophoresis / Temperature Gradient Gel Electrophoresis) analysis method of specific 18S rDNA (Deoxyribose Nucleic Acid) fragment without being based on a GC clamp strategy. The specific 18S rDNA fragment is a fragment containing a terminal region of 18S rDNA, which is far away from an ITS (Internal Transcript Space) sequence; and with a 18S rDNA sequence of a Saccharomyces cerevisiae standard strain as a reference, the terminal region of the 18S rDNA, which is far away from the ITS sequence, is equivalent to a fragment which starts from any nucleotide within a range of the first to the 30th and ends with the 291th-locus nucleotide. According to the invention, a result consistent with that obtained by the use of the GC clamp can be obtained under the same condition by adopting the DGGE / TGGE analysis of the specific 18S rDNA without adopting the GC clamp, therefore, the experiment cost can be reduced and the experiment operation steps are reduced.

Description

technical field [0001] The present invention relates to a DGGE / TGGE analysis method for a specific 18S rDNA fragment, in particular to a DGGE / TGGE analysis method for a specific 18S rDNA fragment not based on the GC splinting strategy. Background technique [0002] DGGE (Denaturing Gradient Gel Electrophoresis) and TGGE (Temperature Gradient Gel Electrophoresis) are two electrophoresis techniques based on the sequence difference of DNA fragments. They were commonly used in the effective detection of gene mutations in the early days and have become an important means for the study of microbial diversity. . These two techniques can separate DNA fragments of the same length but different sequences. The separation of the fragments depends on the different solutions of double-stranded DNA when swimming in a gel containing a chemical denaturant gradient (DGGE) or a temperature gradient (TGGE). chain behavior to achieve. In order to detect all the mutations in the sequence as eff...

AI Technical Summary

Problems solved by technology

However, when studying the structure of the flora, it is necessary to simultaneously synthesize two sets of primers including long primers containing GC splints and common primers without GC splints, especially long primers containing GC splints usually require higher costs above 60 bp, and in addition Occasionally reduced amplification efficiency with GC long primers

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