Fast extraction method of adnascent microbe community total DNA of sponge

A technology of microbial community and sponge symbiosis, applied in the direction of microbial measurement/inspection, biochemical equipment and methods, sugar derivatives, etc., can solve the problems of sponge sample versatility, unsuitability, and cumbersome extraction steps, etc.

Inactive Publication Date: 2006-03-01
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Benefits of technology

The technology described by this inventor allows for quicker identification of bacteria from complex samples like blood or urine that are associated with human body tissues such as joints and muscles. This makes it possible to study how these organism communities work together better over time.

Problems solved by technology

This patented technical problem addressed in this patent relates to developing a quick and effective process for obtaining useful quantities of specific types of biomolecules called ribbon synergism proteins (RSP), particularly those involved in producing certain enzyme activities such as glutathione peroxidase activity.

Method used

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  • Fast extraction method of adnascent microbe community total DNA of sponge
  • Fast extraction method of adnascent microbe community total DNA of sponge
  • Fast extraction method of adnascent microbe community total DNA of sponge

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Embodiment Construction

[0015] The technical solutions of the present invention will be further described below in conjunction with the accompanying drawings and embodiments.

[0016] Realize the technical route of the inventive method as figure 1 As shown, the sponge sample was obtained first, the sponge tissue was mechanically ground and lysed, and the cell wall was broken to release DNA, and the total DNA of the sponge symbiotic microbial community was extracted. The extracted DNA samples can be used for microbial population structure composition and diversity analysis.

[0017] When the present invention is implemented, carry out according to the following steps:

[0018] 1. Mechanical grinding and lysis of spongy tissue

[0019] Take 5 grams of sponge sample and cut it into 2mm under sterile conditions 3 Small pieces, transfer and place in a mortar in an ice bath, add TE buffer solution (pH=8.0) prepared by 10mM Tris and 1mM EDTA, grind directly to break and lyse the sponge tissue, the time i...

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Abstract

The fast extraction method of adnascent microbe community total DNA of sponge includes cutting sponge sample into small blocks, suspending on TE buffering liquid and set on ice for direct grinding to cracking sponge tissue, adding lysozyme, sodium dodecyl sulfonate and proteinase K into the cracked sponge, centrifuging and adding Tris-saturated phenol to extract, extraction with Tris-saturated phenol, chloroform and isopentanol solution for several times, adding NaCl and isopropanol to obtain DNA precipitate, washing and dissolving in TE buffering solution, slaking with RNA enzyme, and final agarose gel electrophoresis analysis to detect extracted DNA sample. The said method can obtain total DNA sample of adnascent microbe community in large segment of sponge for the need of adnascent microbe community structure and diversity analysis based on nucleic acid.

Description

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Claims

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Application Information

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Owner SHANGHAI JIAO TONG UNIV
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