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Method for preparing nucleic acid sample

A nucleic acid and analyte technology, applied in the field of nucleic acid sample preparation, can solve the problem of pain associated with hair removal

Active Publication Date: 2018-10-23
KAO CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the disadvantages of collecting nucleic acid by peeling off the stratum corneum are: a special tape must be used;
In addition, the disadvantages of collecting nucleic acid from plucked hair are: pain associated with plucking; and, only expression information from hair follicles can be obtained from the collected nucleic acid, etc.

Method used

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  • Method for preparing nucleic acid sample
  • Method for preparing nucleic acid sample
  • Method for preparing nucleic acid sample

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preparation example Construction

[0019] (Preparation method of nucleic acid)

[0020] In one aspect, the present invention provides a method for preparing nucleic acid derived from skin cells of a subject. In one embodiment, the method for preparing nucleic acid from skin cells of a subject according to the present invention includes a step of isolating nucleic acid from skin surface lipid collected from the subject.

[0021] "Skin Surface Lipids (SSL)" in this specification refers to fat-soluble components present on the surface of the skin, and may also be referred to as sebum. Generally speaking, SSL mainly consists of secretions secreted by exocrine glands such as sebaceous glands located on the surface of the skin, and the secretions cover the surface of the skin to form a thin layer.

[0022] In this specification, unless otherwise specified, "skin" refers to a general term for tissues including epidermis, dermis, hair follicles, sweat glands, sebaceous glands, and other glands on the body surface.

...

Embodiment 1

[0069] Example 1: Analysis of RNA in various biological samples

[0070] RNA was analyzed in stratum corneum of skin, superficial skin lipid (SSL), serum, urine and saliva.

[0071] According to the method described in Non-patent Document 2, using acrylic tape (2.5cm×3.0cm) from the same part of the horny layer of the subject's forehead, four cuticles were continuously collected along the depth direction, and then RNA was extracted.

[0072] Using an oil-absorbing film (3M Japan Co.), SSL was collected from the entire face of the subject. Next, the oil-absorbing membrane was cut to an appropriate size, and RNA was extracted using Trizol (registered trademark) reagent (Life Technologies Japan Ltd.) according to the attached protocol.

[0073] Human serum, urine and saliva were purchased from cosmobio. The saliva was centrifuged at 15,000 rpm for 5 minutes to remove mucin and oral cavity cells mixed therein, and the supernatant was transferred to a new test tube and used as a ...

Embodiment 2

[0076] Example 2: The proportion of RNA from each species contained in SSL

[0077] In order to calculate the ratio of the RNA from each species contained in the SSL, according to the absolute quantification method using real-time PCR (Real-timePCR), the common rRNA sequences in bacteria, the common rRNA sequences in fungi, and the human rRNA sequences in SSL were respectively analyzed. The copy number of the rRNA sequence is calculated.

[0078] For bacterial 16S rRNA, the 16S rRNA region of Staphylococcus epidermidis JCM2414 strain was cloned into pUC118. For fungal 26S rRNA, the 26S rRNA of Malassezia globosa strain CBS7874 was cloned into pUC118. For human 18S rRNA, the human 18S rRNA region was cloned into pcDNA3.1. These plasmid DNAs were used as standard samples for real-time PCR (Real-time PCR). PCR primers for bacteria are set based on the general sequence of 16S rRNA (Appl Environ Microbiol, 2012, 78, 5938-5941); PCR primers for fungi are based on the general sequ...

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Abstract

Provided is a method with which a nucleic acid sample can be less invasively collected from a subject. The method for preparing a nucleic acid derived from a skin cell of a subject comprises separating a nucleic acid from a lipid, on a skin surface, collected from a subject.

Description

technical field [0001] The invention relates to a method for preparing a nucleic acid sample. Background technique [0002] In recent years, with the rapid development of analytical technology, it has become possible to perform detailed analysis of molecules (nucleic acid, protein, metabolites, etc.) in various biological samples. Furthermore, techniques are being developed to use these molecular analyzes to investigate present and even future physiological states in the human body. Among them, for analysis using nucleic acid molecules, a comprehensive analysis method has been established, which has the advantages that abundant information can be obtained by a single analysis, and research reports based on many single nucleotide polymorphisms or RNA functions can be easily Make functional connections to the analysis results. [0003] Many research institutions around the world are developing technologies for the diagnosis and prediction of diseases, etc., using nucleic aci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806C12N15/09
CPCC12Q1/6883C12Q1/6886C12Q1/6806C12Q2600/178C12Q2600/112C12Q2600/16C12Q2600/166C12Q2600/158C12Q2600/118C12Q2527/125B32B5/18C12N15/1003C12Q1/68C12N15/09C12Q1/686
Inventor 井上高良八谷辉
Owner KAO CORP