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Oligomeric nucleic acid molecule and its application

An oligonucleotide and molecular technology, used in DNA preparation, DNA/RNA fragments, recombinant DNA technology, etc., and can solve problems such as hybridization

Inactive Publication Date: 2014-02-12
杜权
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although the above methods can isolate small RNAs, the products usually obtained are mixed with other non-target RNAs

Method used

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  • Oligomeric nucleic acid molecule and its application
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  • Oligomeric nucleic acid molecule and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Embodiment 1, oligonucleotide synthesis

[0066] The DNA oligonucleotides used in the present invention were synthesized by Invitrogen Beijing Branch, and the RNA oligonucleotides used were synthesized by Guangzhou Ruibo Biotechnology Co., Ltd.

Embodiment 2

[0067]Example 2. Extraction of RNA in tissue samples

[0068] The surgically isolated tissue samples were obtained, the total RNA was extracted with the animal tissue RNA purification kit from NORGEN, and the extracted total RNA was quantified.

Embodiment 3

[0069] Example 3. Separation and RNA Extraction of Serum and Plasma Samples

[0070] Serum and plasma sample separation

[0071] Obtain blood samples from tested individuals and reference healthy individuals, and separate plasma or serum components from them, the specific steps are as follows:

[0072] 1. Separation of serum: The obtained blood sample is left at room temperature to allow it to coagulate naturally, and then it is subjected to centrifugation, and the supernatant obtained is serum. The centrifugation conditions were 4°C, 500×g, and the centrifugation time was 10 minutes.

[0073] 2. Separation of plasma: Anticoagulant treatment is performed on the obtained blood sample according to the techniques known in the art, followed by centrifugation to remove blood cells, and the obtained supernatant is plasma. The centrifugation conditions were 4°C, 500×g, and the centrifugation time was 10 minutes.

[0074] Isolation of RNA components from serum and plasma sample...

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Abstract

The invention provides an oligomeric nucleic acid molecule used for small molecular RNA separation. On the one hand, the oligomeric nucleic acid molecule can be formed by connecting two nucleic acid fragments, wherein a first nucleic acid fragment is a general sequence with a hairpin structure, the 5' end of the sequence is phosphorylated, part of the sequence at the 3' end is complementary with part of the sequence at the 3' end of a second nucleic acid fragment, and the first nucleic acid fragment is connected to a screenable marker; the second nucleic acid fragment includes two nucleic acid sequence regions, the 5' end region is completely or partially complementary with a to-be-separated small molecular RNA sequence, and the 3' end region is completely or partially complementary with part of the sequence at the 3' end of the first fragment; and on the other hand, the oligomeric nucleic acid molecule is a directly synthesized nucleic acid fragment containing the sequence properties. Further, the invention provides a method for separating small molecular RNA by the oligomeric nucleic acid molecule, and a kit using the oligomeric nucleic acid molecule and the method to purify small molecular RNA.

Description

technical field [0001] The present invention relates to the field of biotechnology. More specifically, the present invention relates to an oligomeric nucleic acid molecule used for isolating small molecular RNA, a method for isolating small molecular RNA using the oligomeric nucleic acid molecule and a kit for the method. Background technique [0002] Small RNAs—RNA molecules of 100 nucleotides or less—are currently an area of ​​great interest and promise in the field of molecular therapeutics. These small RNA molecules mainly include microRNA molecules (microRNA, abbreviated as miRNA) and small interfering RNA molecules (small interfering RNA, abbreviated as siRNA). Since these two RNA molecules can hybridize with target mRNA, they can regulate gene expression. In different tissues, organs and different developmental stages, the expression of miRNA and / or siRNA is different, and they may play an important regulatory role in various biological development, physiological act...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/10
Inventor 杜权
Owner 杜权
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