Kit for detecting micro ribonucleic acid in blood serum and preparation method thereof
A technology of microRNA and its production method, which is applied in the biological field, can solve the problems of lack of microRNA kits, etc., and achieve the effect of simple and clear results, accurate results, and low cost
Active Publication Date: 2014-06-25
JIANGSU MICROMEDMARK BIOTECH
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However, because the research work on human and animal microRNA has only been carried out for about 4 years, there is no relevant microRNA kit at present.
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[0034] 1. RT-PCR experiment of microRNA in serum
[0035]We used RT-PCR technology to discover and prove for the first time that various microRNAs can be detected in human and animal serum, and their expression is quite abundant. The specific steps are: (1) Collect the serum of mice, rats, normal people and some patients. (2) Preparation of cDNA samples. There are two schemes for this operation. One scheme is to use 10 μl of serum as a buffer to directly perform reverse transcription reaction, and the other is to use Trizol reagent (Invitrogen) to extract serum total RNA first (10ml serum can usually enrich about 10 μg of RNA) , and then obtain cDNA by RNA reverse transcription reaction. The reverse transcription reaction system includes 4 μl 5×AMVbuffer, 2 μl 10mM each dNTP mixture (Takara Company), 0.5 μl RNase Inhibitor (Takara Company), 2 μl AMV (Takara Company) and 1.5 μl gene-specific reverse primer mixture, and the reaction step is 16 Incubate at ℃ for 15 minutes, re...
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The invention belongs to the technical field of biology and relates to a kit for detecting micro ribonucleic acid in blood serum and a preparation method thereof. The kit disclosed by the invention is prepared by a method comprising the following steps: I, determining that the normal ribonucleic acid contains more than one copied micro ribonucleic acid by a sequencing method or a quantitative PCR (Polymerase Chain Reaction) method; II, screening one type of micro ribonucleic acid which has a large expression amount and a great difference degree in disease and normal physiological states through a biochip technology, wherein the type of micro ribonucleic acid is used as a probe for predicating whether canceration or other diseases happen and diagnosing the pathological change degree; and III, preparing the type of micro ribonucleic acid probe prepared in the step II and reagents containing Taq enzyme and dNTP into the kit for detecting the micro ribonucleic acid in the blood serum. The kit is put into practice and particularly increases the possibility of finding pathological changes of diseases, which are not suitable for biopsy examination, at an early stage, can help and guide diagnosis and treatment and can push definite diagnosis and treatment of the diseases to a new height.
Description
[0001] Case Description [0002] This case is the application number 200710134620.4, the application date is November 2, 2007, and the name is the detection method of microRNA in serum, the kit used for detection, biochip and its production and application method. case application. technical field [0003] The invention belongs to the field of biotechnology, and in particular relates to a kit for detecting microRNA in serum and a preparation method thereof. technical background [0004] Micro ribonucleic acid, English name microRNA, abbreviated as miRNA, is a kind of non-coding single-stranded small ribonucleic acid molecules with a length of about 19 to 23 nucleotides. They are highly conservative in evolution and widely exist in animal and plant cells. In the past five years, hundreds of microRNA molecules have been identified in various biological species such as humans, mice, and rats. MicroRNA inhibits the translation of target mRNA by recognizing the 3' untranslated ...
Claims
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IPC IPC(8): C12Q1/68
Inventor 张辰宇张峻峰陈熹巴一陈江宁王可慧殷媛蔡星张燕张红杰曾柯王进郭继刚
Owner JIANGSU MICROMEDMARK BIOTECH