A method for extracting high-quality total RNA from grass carp mesenteric adipose tissue
A technology of adipose tissue and extraction method, applied in the direction of recombinant DNA technology, DNA preparation, etc., can solve the problem of not being able to stably obtain high-quality grass carp adipose tissue RNA, and achieve the reduction of RNA degradation, stable effect, purity and integrity high effect
Active Publication Date: 2022-03-04
PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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The test also found that the currently published optimization and improvement methods or strategies failed to stably obtain high-quality grass carp adipose tissue RNA
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[0035] The present invention will be further described in detail below in conjunction with examples, but the embodiments of the present invention are not limited thereto. Unless otherwise specified, the reagents, equipment and methods used in the present invention are commercially available reagents, equipment and routinely used methods in this technical field.
[0036] 1. Materials and methods
[0037] 1.1 Test fish
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The invention discloses a method for extracting high-quality total RNA from grass carp mesenteric adipose tissue. % ethanol washing and other steps. In the sample pretreatment stage, the present invention can increase the yield to meet the demand for RNA quantity in conventional molecular biology experiments by increasing the sampling amount to 30mg; Long-term mechanical homogenization, 3 minutes long, can significantly reduce the occurrence of RNA degradation, and at the same time add 2 centrifugation to remove the oil layer and other operations, the purity and integrity of the total RNA obtained are higher. The extraction method of the present invention effectively solves the problems of easy degradation and low yield of grass carp adipose tissue RNA extraction, and has simple operation and stable batch processing effect, which can meet the requirements of high-quality RNA such as transcriptome sequencing, and is also applicable to other fish fats Extraction of total RNA from tissues.
Description
technical field [0001] The invention relates to the field of fish RNA extraction, in particular to a method for extracting high-quality total RNA from grass carp mesentery adipose tissue. Background technique [0002] Animal adipose tissue is not only the body's energy storage, but also an active endocrine organ, which secretes a variety of hormones and cytokines that play an important role in the body's metabolic balance and immune response (Scherer, 2006; Wellen, Hotamisligil, 2003). In the study of tissue function, research at the molecular level is essential, and RNA extraction and purification are often involved in molecular biology experimental techniques. However, because adipose tissue is rich in oil and low in RNA abundance, it is often difficult to extract high-quality total RNA, which affects the efficient development of downstream experiments. [0003] At present, the classic acidic phenol-guanidine isothiocyanate-chloroform one-step extraction method is widely ...
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IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 姜鹏李胜杰樊佳佳马冬梅杜金星
Owner PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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