Kit and extraction method for extracting micro ribonucleic acid (RNA)

A kit and tiny technology, which is applied in the kit for extracting microRNA and its extraction field, can solve the problems of poor efficiency of miRNAs, achieve the effect of enhancing the magnetic field and improving the acquisition rate

Active Publication Date: 2014-05-14
SHANGHAI DIDA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the efficiency of extracting miRNAs using this kit is poor

Method used

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  • Kit and extraction method for extracting micro ribonucleic acid (RNA)
  • Kit and extraction method for extracting micro ribonucleic acid (RNA)
  • Kit and extraction method for extracting micro ribonucleic acid (RNA)

Examples

Experimental program
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Effect test

Embodiment 1

[0041] Please combine figure 1 In an embodiment of the present invention, a kit for extracting microRNA is provided, comprising a box body 101, a reagent bottle 102 containing a first magnetic bead liquid, a reagent bottle 103 containing a second magnetic bead liquid, and a reagent bottle 103 containing a second magnetic bead liquid. The reagent bottle 104 of lysate, the reagent bottle 105 of eluent and the reagent bottle 106 of washing liquid are housed;

[0042] A foam pad 107 is provided on the inner bottom wall of the box body 101; and the edge of the foam pad 107 is attached to the inner side wall of the box body 101;

[0043] Foam pad 107 is provided with five grooves, the reagent bottle 102 that the first magnetic bead liquid is housed, the reagent bottle 103 that the second magnetic bead liquid is housed, the reagent bottle 104 that lysate is housed, the reagent bottle 104 that eluent is housed. The reagent bottle 105 and the reagent bottle 106 containing the cleaning s...

Embodiment 2

[0049] In this embodiment, in order to arrange foam pads 107 and a plurality of reagent bottles in the box body 101 for convenience: preferably, the box body 101 is a rectangular box body with an opening, and the opening of the rectangular box body is opposite to the bottom wall; all The grooves all extend from the bottom wall of the rectangular box to the opening.

[0050] In addition, in order to facilitate the reagent bottle to be placed on its corresponding groove, it is also to prevent the reagent bottle from being easily separated from the groove after the box body 101 falls down during transportation, thereby causing unexpected phenomena such as reagent bottle breakage, Preferably, all the reagent bottles are cylindrical, and the cross-sections of all the grooves are circular; the inner wall of each groove fits the outer wall of the corresponding reagent bottle.

[0051] In addition, in this embodiment, in order to increase the specific surface area of ​​the first magne...

Embodiment 3

[0054] This embodiment specifically includes the following steps:

[0055] Step 101: adding a lysis solution to the serum or plasma sample for lysis to obtain a lysis mixture, centrifuging the lysis mixture to obtain a supernatant;

[0056] This step is the process of lysing the sample. Through the added lysate, the encapsulated nucleic acid in the sample is freed. After centrifugation, large particles of impurities are precipitated, and microRNA is contained in the supernatant for subsequent processing. separation.

[0057] In addition, in the above steps, in order to improve the lysis efficiency and release the microRNA in the sample as completely as possible, preferably, the lysate includes surfactants, isothiocyanates, guanidinium salts, lithium salts, RNase inhibitors reagents, buffers with a pH of 6.5-8.5, or a combination of these reagents.

[0058] Moreover, the preferred lysis time is: 10-20 minutes; the centrifugation speed is 10000-12000 rpm; the centrifugation ti...

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Abstract

The invention relates to the technical field of organisms, and particularly relates to a kit and an extraction method for extracting micro ribonucleic acid (RNA). The kit comprises a box body, a reagent bottle containing first magnetic bead liquid, a reagent bottle containing second magnetic bead liquid, a reagent bottle containing lysate, a reagent bottle containing an eluent, and a reagent bottle containing a scrubbing solution, wherein a foam pad is arranged on the inner bottom wall of the box body; the edge of the foam pad is attached to the inner lateral wall of the box body; five grooves are arranged on the foam pad; one reagent bottle is respectively arranged inside each groove; the first magnetic bead liquid contains a plurality of first magnetic beads, and a targeting sequence or a random sequence capable of being paired with the micro RNA is modified on the surface of each magnetic bead; the second magnetic bead liquid contains a plurality of second magnetic beads, and a hydrophilic high-molecular polymer with negative charge is modified on the surface of each magnetic bead. By adopting the kit disclosed by the invention, the micro RNA in plasma or serum can be more efficiently extracted, and the kit is especially applicable to extraction of the micro RNA with low content or low copy number in a plasma or serum sample.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit for extracting microRNA and an extraction method thereof. Background technique [0002] MicroRNAs (miRNAs) are a class of non-coding endogenous small single-stranded RNAs with a length of 21-25 ribonucleotides. In 1993, Lee et al. first discovered the first miRNA (lin-4) in nematodes, which can regulate the larval development of nematodes by inhibiting the expression of nucleoprotein lin-14 at the translational level. Subsequently, scientists applied random cloning and sequencing, and bioinformatics prediction methods, and found that miRNAs widely exist in primates, rodents, birds, fish, flies, worms, plants, and viruses. From more than 200 species in 2002 to nearly 9,000 species at the end of 2008, the discovery of miRNAs has shown explosive growth, and miRNAs in new species have been discovered continuously. [0003] Human miRNAs genes only account for about 2%-3% of the to...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/00C12N15/10
CPCC12N15/1013
Inventor 陈静葛阳
Owner SHANGHAI DIDA BIOTECH
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