Shredder for mechanical disruption by gentle controlled compressive rotation

A technology of rotating elements and rotating rates, applied in the preparation of test samples, analytical materials, biochemical equipment and methods, etc., can solve the problems of incomplete splitting, unpredictability, unreliable splitting technology, etc., to achieve high productivity, high The effect of productivity

Inactive Publication Date: 2011-09-28
PRESSURE BIOSCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these splitting techniques may be unreliable or even unpredictable, e.g. with respect to splitting levels
For example, if low or insufficient force or energy is applied, fragmentation is typically incomplete and the effective productivity of analyzable molecules is low; whereas if very high or excessive force or energy is applied, high shear Stress and heat will mechanically and thermally (or both) alter the target extraction product and undesirably alter its properties, resulting in a sample that converts to a component that is no longer of interest because the product is less important than the target material not representative

Method used

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  • Shredder for mechanical disruption by gentle controlled compressive rotation
  • Shredder for mechanical disruption by gentle controlled compressive rotation
  • Shredder for mechanical disruption by gentle controlled compressive rotation

Examples

Experimental program
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Effect test

example 1

[0057] Example 1. In physiological buffer or ProteoSOLVE TM In the case of IEF reagent, the pulverizer and BIOMASHER of the present invention when extracting protein from pine needles TM (From Nippi, Inc.) Comparison.

[0058] The pine needles are roughly cut to about 4-5mm length within one hour of harvest. 50 or 200mg is weighed to the tared PULSE Tubes or BioMasher TM Inset. Samples are in duplicate in KPO 4 Buffer or ProteoSOLVE IEF reagent with 100mM DTT.

[0059] As in Image 6 As shown in, for BioMasher TM In the centrifugal method, the components are centrifuged at 14,000 for 20 seconds under the condition that the homogenizer rod is positioned according to the manufacturer's instructions. BioMasher TM The insert is 80-140um micropore size. The insert was washed twice with 700 μL each time and centrifuged. Combine the initial homogenized product and washings. The final sample volume is 1400 μL. For BioMasher TM Rotating the grinding method, connect the homogenize...

example 2

[0076] Example 2. Using PCT Shredder TM And Pressure Cycle Technology (PCT) to increase protein productivity from coniferous plants

[0077] Plant proteins (proteome) provide an opportunity to monitor post-translational responses to environmental impacts such as pollution, pests, or plant diseases. Comprehensive protein analysis requires reliable extraction methods that can separate proteins reproducibly and without bias. The sample preparation of plant tissues is particularly complicated due to the fact that the nature of the cell wall makes it difficult to extract analytes quantitatively; the relatively low protein cell content in some plant tissues; or the abundance of lignin, tannic acid, and other polyphenols, This can interfere with protein analysis. The extraction of protein from pine needles or other coniferous tissues is particularly complicated and may be further complicated by their high content of terpene resin in these species. Here is a description of a system fo...

example 3

[0101] Example 3. Analysis of Tick Borrelia and HGE gene expression of DNA preparations separated by pulverizer and PCT: standard curve and total bacterial decomposition.

[0102] The basic method used for tick DNA extraction involves the following steps.

[0103] Before PCT, soak the tick sample in Tris buffer for 1 hour. A tick was loaded into the end of the punch and crushed by hand, followed by 60 cycles of PCT treatment in proteinase K at 56C. Place the test tube in boiling water and boil for 10 minutes, then unload. The CTAB buffer was added to a final concentration of 2% and allowed to incubate at 65C for 20 minutes. Carry out phenol-chloroform purification. Store the final volume of 100ul at -20C.

[0104] Perform real-time PCR. For the relative amounts of Borrelia and total bacterial DNA, two standard curves were designed. In doing so, the Borrelia DNA and E.coli DNA from ATCC were diluted in stages. Amplify the Borrelia 23S rRNA gene and the bacterial 16SrDBA gene.

...

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Abstract

The systems and techniques of the present invention can also synergistically utilize mechanical disruption processes with the use of high hydrostatic pressure extraction, such as pressure cycling extraction techniques to achieve high yield of difficult to extract sample constituents without generating high shear stress or high temperatures.

Description

[0001] Cross reference to related applications [0002] This application claims the right of priority to U.S. Patent Application No. 6I / 097,830 filed on September 17, 2009, entitled SHREDDER FOR MECHANICAL DISRUPTION BY GENTLE CONTROLLED COMPRESSIVE ROTATION, the entire content of which is incorporated by reference for all purposes Included here. Technical field [0003] The present invention relates to the provision and preparation of samples for analysis. In particular, the present invention relates to the preparation of biological samples so as to promote small molecules (such as deoxyribonucleic acid) by crushing the biological samples under the splitting force generated by the force caused by rotation. , Ribonucleic acid, lipid, protein) extraction and analysis. Background technique [0004] When biological samples have tough external structures, it is very difficult to extract DNA, RNA, proteins, lipids, and small molecules from these samples. For many sample types, if their...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28
CPCC12M35/04G01N1/286G01N2001/2866
Inventor E·Y·廷A·拉扎雷夫V·格罗斯C·杜莎尔特C·李N·劳伦斯R·T·舒马赫
Owner PRESSURE BIOSCI
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