Kit and method for rapidly releasing nucleic acid from paraffin-embedded tissue sections

Abstract

The invention discloses a kit and a method for rapidly releasing nucleic acid from paraffin-embedded tissue sections. The kit comprises a proteinase K solution (prepared from 10-100g / L protease K, 10-200mM / L Tris with pH value of 7.0-11.0 and 0.01-5mM / L anhydrous calcium chloride), a first nucleic acid release reagent (1-1000 mM / L Tris with pH value of 7.0-11.0, Triton X-100 with a volume ratio of0.1 to 2%, Nonidet P-40 or substitutes thereof with a volume ratio of 0.1 to 2%, and 0.1-10mM / L EDTA disodium salt), and a second nucleic acid release reagent (10-200mM / L Tris with pH value of 7.0-8.0, and Nonidet P-40 or substitutes thereof with a volume ratio of 0.1- 2%). The kit is also preferentially provided with a centrifuge column; when the kit is in use, the paraffin section is placed ina centrifuge tube, the first nucleic acid release reagent and the proteinase K solution are added into the centrifuge tube, the materials in the centrifuge tube are fully and evenly mixed, and incubation is carried out for the first time; after that, the second nucleic acid release reagent is added and is evenly mixed in a way of vortex, and incubation is carried out for the second time, so that asplit product is obtained; the product is then filtered by means of the centrifuge column, so that target nucleic acid can be obtained. After the kit and the method are adopted, the paraffin sectionsdo not need to be subjected to dewaxing, and the complicated purification step is omitted. The kit and the method have the advantages of being safe to use, simple and convenient to operate, high in recovery efficiency and lower in manufacturing cost.

Description

technical field [0001] The invention relates to the field of nucleic acid extraction in biotechnology, in particular to a kit and method for rapidly releasing nucleic acid from paraffin-embedded tissue sections. Background technique [0002] The main methods of nucleic acid extraction include: (1) organic reagent extraction method, (2) spin column method, (3) magnetic bead method, in which: [0003] (1) Organic reagent extraction method, the so-called phenol-chloroform extraction method. Because it is easier to remove the protein by alternating extraction with two different organic solvents, the DNA extracted by this method has high purity, large fragments, and good results. Use of self-prepared reagents in the laboratory. [0004] (2) Spin column method, in the presence of high-concentration chaotropic salt, nucleic acid components released from cells (including bacteria) or virus particles can be bound to silica, and then purified nucleic acid can be obtained through was...

AI Technical Summary

Problems solved by technology

[0006] The archives of paraffin tissues represent precious and widely sourced materials for biomedical research. However, due to the fixation of formaldehyde solution (formalin) in the preparation process, the DNA molecules in the sample will be broken and cross-linked. Extraction with organic reagents, centrifugation The column method will cause the loss of nucleic acid fragments in the purification process of repeated operations, and the recovery efficiency is low

Images