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Extraction method for total RNA from secondary follicle tissue of Cashmere goat

A technology of secondary hair follicles and cashmere goats, which is applied in the field of total RNA extraction, can solve the problems of difficult grinding, troublesome sampling work, RNA yield and quality can not be guaranteed, and achieve the effect of good integrity and high purity

Inactive Publication Date: 2014-08-13
YANCHENG TEACHERS UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the small size of cashmere goat secondary hair follicle tissue, it is difficult to grind after freezing in liquid nitrogen and floating on the surface of liquid nitrogen, and the RNA content of secondary hair follicle tissue is relatively low, so the yield and quality of RNA extracted by this method cannot be guaranteed
Especially under field conditions, carrying liquid nitrogen tanks to store and transport hair follicle tissue samples will also be subject to certain restrictions, which will bring great trouble to the sampling work

Method used

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  • Extraction method for total RNA from secondary follicle tissue of Cashmere goat
  • Extraction method for total RNA from secondary follicle tissue of Cashmere goat
  • Extraction method for total RNA from secondary follicle tissue of Cashmere goat

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] (1) Preparation before extraction of total RNA from secondary hair follicle tissue

[0035] The total RNA of secondary hair follicle tissues of different cashmere goats was extracted by silica gel membrane adsorption column method. The extraction process is as follows:

[0036] ①Sample collection and processing

[0037] The secondary hair follicle tissues of 5 Shanbei white cashmere goats were collected from a farm in the suburbs of Yulin City, Shaanxi Province. The side of each sheep was wiped and sterilized with iodine tincture and alcohol cotton balls, and 5 small pinches of wool and cashmere mixture were extracted and put into 1.5mL RNase-free finger tubes filled with RNAlater preservation solution to obtain 5 The wool and cashmere mixture samples were tightly capped and placed in an incubator with frozen ice packs, transported back to the laboratory (the time from sampling to delivery to the laboratory was 4 days), and stored in a 4°C refrigerator.

[0038] Each...

Embodiment 2

[0042] Example 2 Extraction of total RNA from the secondary hair follicle tissue of sample 1

[0043] The specific extraction process of the total RNA of the first secondary hair follicle tissue sample is as follows:

[0044] 1) Quickly transfer the isolated secondary hair follicle tissue to the lysate (the lysate is pH 7.0, the concentration is 30mmol / L Tris-HCl, 5mol / L guanidine isothiocyanate, and 1% β-mercaptoethanol into), use a grinding pestle to grind the secondary hair follicle tissue in the lysate, fully grind to break the tissue to obtain a mixed solution, the grinding time is 5min, carefully remove the hair follicle with pointed tweezers; because the hair follicle is attached to the hair follicle, grind After the tissue falls off, the velvet stem needs to be removed.

[0045] 2) Use a pipette to transfer the ground mixture to a finger tube, centrifuge at 10,000rpm for 3min, and carefully transfer the supernatant to a 1.5mL RNase-free finger tube;

[0046] 3) Add a...

Embodiment 3

[0052] Example 3 Extraction of total RNA from the secondary hair follicle tissue of sample 2

[0053] The specific extraction process of the total RNA of the second secondary hair follicle tissue sample is as follows:

[0054] 1) Quickly transfer the isolated secondary hair follicle tissue to the lysate (the lysate is pH 7.0, the concentration is 30mmol / L Tris-HCl, 5mol / L guanidine isothiocyanate, and 1% β-mercaptoethanol into), use a grinding pestle to grind the secondary hair follicle tissue in the lysate, fully grind to break the tissue to obtain a mixed solution, the grinding time is 10min, carefully remove the hair follicle with pointed tweezers; because the hair follicle is attached to the hair follicle, grind After the tissue falls off, the velvet stem needs to be removed.

[0055] 2) Use a pipette to transfer the ground mixture to a finger tube, centrifuge at 10,000rpm for 5min, and carefully transfer the supernatant to a 1.5mL RNase-free finger tube;

[0056] 3) Add...

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Abstract

The invention discloses an extraction method for high-quality total RNA from the secondary follicle tissue of a Cashmere goat, which relates to the field of molecular biology. The extraction method comprises the following main steps: acquisition, preservation and transport of the secondary follicle tissue; separation of the secondary follicle tissue; extraction of total RNA of the secondary follicle tissue; and detection of quality of a total RNA sample. The method is simple to operate, has good stability and is suitable for common laboratory operation. The extracted total RNA of the secondary follicle tissue has high purity and good integrity and can be further applied to scientific research in related fields like preparation of cDNA, cloning of a target gene, real time fluorescent quantitative PCR detection and high flux sequencing. The method is also applicable to extraction of total RNA from follicle tissue of other animals.

Description

technical field [0001] The invention relates to the field of animal molecular biology, in particular to a method for extracting total RNA from secondary hair follicle tissues of cashmere goats. Background technique [0002] Cashmere is the main product of cashmere goats. It is a precious textile raw material with extremely high economic value. The down is a derivative of the skin of the cashmere goat. The primary hair follicles grow coarse hairs, and the secondary hair follicles grow down. The secondary hair follicle of cashmere goats is a precise regenerative organ, and its periodic changes not only control the growth and development of down hair, but also are closely related to the amount of cashmere produced and the quality of down hair. The periodic change process of secondary hair follicles is a molecularly regulated process, which is realized through the interaction of a series of signaling pathways and genes. In terms of the molecular regulation mechanism of the per...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 耿荣庆王兰萍
Owner YANCHENG TEACHERS UNIV
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