Multifunctional constant-speed heating-denatured gradient gel electrophoresis system

A technology of denaturing gradient gel and electrophoresis system, applied in the field of DNA electrophoresis, which can solve the problems of reducing resolution and affecting gel separation effect, and achieve the effect of improving gel resolution and shortening electrophoresis time

Inactive Publication Date: 2013-02-13
谭啸
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  • Abstract
  • Description
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Problems solved by technology

Both DGGE and TGGE use polyacrylamide gel with constant monomer concentration (ie, the gel pore size is consistent), which affects the separation effect of the gel and reduces the resolution (Cremonesi et al., 1997; Petri & Imhoff, 2001)

Method used

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  • Multifunctional constant-speed heating-denatured gradient gel electrophoresis system

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Embodiment 2

[0033] Set the preheating time = 1h, initial temperature = 60°C, heating time = 0h, final temperature = 60°C, holding time = 16h, gel denaturant concentration gradient of 40% to 50%, the total DNA of 13 lake water samples PCR products were subjected to DGGE.

[0034] As shown in the result figure, the TTGG electrophoretic spectrum of the present invention ( figure 1 A) The bands are clear and the resolution is high; the DGGE electrophoresis pattern ( figure 1 B) Bands are fuzzy and low resolution.

[0035]In terms of electrophoresis time, TTGG (7 hours in total) is more than half shorter than DGGE (17 hours in total). For the same 13 samples, the resolution of the TTGG map is higher than that of the DGGE map, and the single-lane band diversity (Shannon-Weiner index, H’ ) increased by more than 10%. Single lane band diversity index ( H’ ) is calculated as follows:

[0036] H’ = -Σ (Pi) (lnPi)

[0037] Pi (the relative gray value of a certain band) = ni / N, ni: the gray ...

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Abstract

The invention relates to a multifunctional constant-speed heating-denatured gradient gel electrophoresis system which is capable of overlapping a gel gradient on a space in a temporal heating process so as to reduce the electrophoresis time and increase the gel resolution. According to the electrophoresis system provided by the invention, constant-speed heating-denatured gradient gel electrophoresis, denatured gradient gel electrophoresis and common polyacrylamide gel electrophoresis can be carried out, and a constant-temperature cold groove is externally connected so as to preform single strand conformation polymorphism, so that one machine has multiple purposes. According to the electrophoresis system, microbial diversity and the relative content of the microbes are quickly and effectively revealed, the functional flora is searched and harmful flora is prevented, and the electrophoresis system has a huge action on industry, agriculture, medicine, environmental monitoring and pollution abatement.

Description

technical field [0001] The invention relates to the field of DNA electrophoresis, in particular to a uniform heating-denaturing gradient gel multifunctional electrophoresis system. Background technique [0002] Microbial communities are structural and functional units that widely exist in ecosystems. Different microorganisms in the community co-exist in an orderly manner, and they have their own nutritional pathways and metabolic types. Microbial community structure research is a hotspot in ecology and environmental science research, involving soil and rhizosphere microorganisms, intestinal microorganisms, planktonic microorganisms and other fields. For the development of microbial resources, it clarifies the relationship between microbial communities and habitats, and reveals community structures and functions. It is of great significance to guide the directional regulation of microbial community functions. [0003] Microbial community structure analysis technology is divi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/447
Inventor 谭啸
Owner 谭啸
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