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Improved method for industrial-scale manufacturing of DNA

An industrial-scale, clear liquid technology, applied in the field of manufacturing high-purity DNA, can solve the problems of high difficulty and unseen product supply, and achieve the effect of simple operation

Active Publication Date: 2014-06-18
NANTONG QIUZHIYOU BIOSCI & BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the preparation of calf thymus is relatively difficult, so there is no product supply

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 DNA Extraction from Calf Thymus

[0022] Take 100Kg of frozen calf thymus and thaw it for a while (it can’t be completely thawed, just thaw until it becomes soft), put it in a meat grinder and grind it twice, add 1000L buffer solution (containing 0.14mol / L sodium chloride, 0.05mol / L sodium citrate buffer, pH7.0), stir evenly, and filter the calf thymus solution with a 20-mesh stainless steel mesh to remove connective tissue. Then use a tube-type high-speed centrifuge to centrifuge, and then wash the precipitate obtained with buffer solution, and repeat the centrifugation twice.

[0023] Put the precipitate in a meat grinder and grind twice, add 800L of buffer solution (containing 0.14mol / L sodium chloride, 0.05mol / L sodium citrate buffer, pH7.0); then put it in a 3000L extraction tank Add buffer solution to make the total volume 1400L, start stirring at 100 rpm, and slowly add 6.3Kg of sodium dodecylsulfonate with a weight content of 5%, stir at room tempera...

Embodiment 2

[0026] Example 2 DNA Extraction from Fish Sperm

[0027] Take 10Kg of frozen salmon sperm (milt) to melt for a while, put it into a meat grinder and grind it, add 100L of buffer solution (containing 0.14mol / L sodium chloride, 0.05mol / L sodium citrate buffer, pH=7.0) Stir evenly, filter with a 40-mesh nylon mesh to remove connective tissue, and then centrifuge with a tubular high-speed centrifuge.

[0028] The following steps are the same as in Example 1 (because the amount of raw materials is 1 / 10 of that of Experimental Example 1, the amount of reagent added is also 1 / 10 of that of Example 1).

[0029] Finally, 321gDNA can be obtained, with a content of 91%, A 260 / A 280 =1.85.

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PUM

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Abstract

The invention provides an improved method for industrial scale manufacturing of DNA, and the improved method comprises the following steps: dissolving sodium dodecyl sulfate SDS in a dissociated DNA solution, adjusting the pH to be alkaline, standing for 1 to 2 hours, centrifuging and collecting a supernatant liquid, adjusting the supernatant liquid pH to 7.0-7.5, adding alcohol for precipitation, collecting precipitation, washing and drying to obtain the DNA product. By adopting the method of the invention, high-content DNA can be obtained, and the method has the advantages of simple operation and is suitable for industrial production.

Description

Technical field: [0001] The invention relates to a method for producing high-purity DNA. Background technique: [0002] Calf thymus, fish sperm and embryos of plant seeds are rich in DNA and are good materials for extracting DNA. It has also been reported to start from the cultivation of microorganisms and then extract DNA from microorganisms, but it has not been used in industrial production. Therefore, the method of extracting DNA from calf thymus or fish sperm (milt) is currently the main method for producing DNA on an industrial scale. [0003] The general preparation method is to firstly remove blood and connective tissue from fresh (or frozen) calf thymus or milt, cut into small pieces in an ice bath, wash with buffer and use a tissue masher (8000 rpm-12000 RPM) mashed, washed with citrate buffer containing 0.14mol / L sodium chloride for several times to remove ribonucleoproteins, and then obtain deoxyribonucleoproteins (or nuclei). In industrial preparation, at leas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 邱蔚然周洁邱志云祁妤琳
Owner NANTONG QIUZHIYOU BIOSCI & BIOTECH