Extraction method of messenger RNA
An extraction method and messenger technology, applied in the biological field, achieves a high success rate and a stable and reliable construction method
Inactive Publication Date: 2019-12-17
XIANGTAN ZHILIAN TECH MATASTASIS PROMOTE CO LTD
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- Application Information
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Problems solved by technology
Various genetic and surgical approaches have been used to attempt to explain the complexity of the neocortex and the differently characterized neuronal cell types, however labeling and purifying and defining neuronal subtypes remains challenging because In , specific markers are always dynamically changing
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Embodiment Construction
[0045] The method for extracting messenger RNA provided by the present invention will be further described in detail and completely below in conjunction with the examples. The embodiments described below are exemplary only for explaining the present invention and should not be construed as limiting the present invention.
[0046] The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples were purchased from the market unless otherwise specified.
[0047] The instruments and reagents adopted in this embodiment are as follows:
[0048] Neuro2a cell line, purchased from ATCC;
[0049] CD-1 mice were purchased from Charles River Laboratories, USA;
[0050] Penicillin-streptomycin, purchased from gibco;
[0051] PolyJet TM DNA In Vitro Cell Transfection Kit, purchased from SignaGen Laboratories
[0052] BCA protein quantification kit was purchased from Thermo Scientific;
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Abstract
The invention discloses an extraction method of messenger RNA. According to the extraction method, plasmids of marked ribosome are transfected into an embryo through an intrauterine embryo electrotransfection technology; and after the plasmids grow for a specific time, a translated ribosome affinity purification technology is performed to obtain purified messenger RNA. According to the invention,the intrauterine embryo electrotransfection technology and the translated ribosome affinity purification technology are combined. Specific neurons are marked at specific time and in a brain area according to time characteristics of neuron development in different lamellar layers of cerebral cortex, and experiments are performed at different time points in the development process, so that a development period dynamic molecular library is expected to be obtained to construct a translation spectrum, and a solid foundation is laid for summarizing rules, discovering specific key factors and screening effective genes.
Description
technical field [0001] The invention relates to a method for extracting messenger RNA, which belongs to the field of biotechnology. Background technique [0002] The mammalian neocortex is a complex and highly organized structure consisting of 6 layers of different types, with hundreds of different types of neuron cells and glial cells in between. The neocortex, the brain region responsible for cognitive function, sensory perception, and consciousness, has undergone significant expansion and development during evolution. During development, each lamella has a fixed developmental sequence and time, and produces characteristic neurons of different lamina in chronological order. Neuron development is based on the central dogma, the complex orchestration of genetic material through transcription and translation, and finally the production of proteins to play biological roles, to obtain specific cellular identities. The temporal and spatial complexity of neuronal development po...
Claims
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Login to View More IPC IPC(8): C12Q1/6806C12N15/85C12N15/65
CPCC12Q1/6806C12N15/85C12N15/65C12N2800/107C12Q2523/308
Inventor 不公告发明人
Owner XIANGTAN ZHILIAN TECH MATASTASIS PROMOTE CO LTD