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Method for extracting DNA from breast cancer paraffin-embedded tissue

A paraffin-embedded, breast cancer technology, applied in the field of DNA extraction, can solve the problems of complex operation steps, large sample loss, increased cross-contamination, etc., and achieves the effects of high DNA yield and purity, simple operation and simple equipment.

Inactive Publication Date: 2016-05-11
TIANJIN YINGSHIBO TECH DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The traditional phenol / chloroform extraction method is the earliest method to purify DNA from paraffin-embedded tissues of breast cancer. Although it can meet the analysis requirements such as PCR, the reagents used are highly toxic, the operation steps are complicated, the sample loss is large, and cross-contamination is also difficult. It is not suitable for a large number of clinical specimens, especially the needs of routine testing

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  • Method for extracting DNA from breast cancer paraffin-embedded tissue
  • Method for extracting DNA from breast cancer paraffin-embedded tissue

Examples

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Effect test

Embodiment 1

[0023] A method for extracting DNA from paraffin-embedded tissues of breast cancer, comprising the steps of:

[0024] The first step: sampling, the sample storage time is 24 hours

[0025] Step 2: Isolate and extract DNA

[0026] ① Dewaxing: Cut the paraffin-embedded tissue to be treated into tissue slices with a thickness of 5 μm, take 3 tissue slices and place them in a sterile 1.5ml centrifuge tube, add 125 μL of extraction buffer with a pH of 8.0, seal them with tape, Put it in a microwave oven and heat it on high heat for 1 minute, take it out and centrifuge it quickly, the centrifugation speed is 14000r / min, and the centrifugation time is 8 minutes, take it out and cool it at -20°C until the water phase and the wax layer organic phase are separated, so that the upper layer of the centrifuge tube The wax sheet is fully hardened, tilt the centrifuge tube, and use a sterilized syringe to suck out the tissue mixture solution containing DNA at the bottom to obtain sample A; ...

Embodiment 2

[0031] A method for extracting DNA from paraffin-embedded tissues of breast cancer, comprising the steps of:

[0032] The first step: sampling, the sample storage time is 10 years

[0033] Step 2: Isolate and extract DNA

[0034] ① Dewaxing: Cut the paraffin-embedded tissue to be treated into tissue slices with a thickness of 10 μm, take 3 tissue slices and place them in a sterile 1.5ml centrifuge tube, add 250 μL of extraction buffer with a pH of 8.5, seal them with tape, Put it in a microwave oven and heat it on high heat for 2 minutes, take it out and centrifuge it quickly, the centrifugation speed is 20000r / min, and the centrifugation time is 12 minutes, take it out and cool it at -20°C until the water phase and the wax layer organic phase are separated, so that the upper layer of the centrifuge tube The wax sheet is fully hardened, tilt the centrifuge tube, and use a sterilized syringe to suck out the tissue mixture solution containing DNA at the bottom to obtain sample ...

Embodiment 3

[0039] A method for extracting DNA from paraffin-embedded tissues of breast cancer, comprising the steps of:

[0040] The first step: sampling, the sample storage time is 8 years

[0041]Step 2: Isolate and extract DNA

[0042] ① Dewaxing: Cut the paraffin-embedded tissue to be treated into tissue slices with a thickness of 8 μm, take 3 tissue slices and place them in a sterile 1.5ml centrifuge tube, add 200 μL of extraction buffer with a pH of 8.3, seal them with tape, Put it in a microwave oven and heat it on high heat for 1 minute, take it out and centrifuge it quickly, the centrifugal speed is 16000r / min, and the centrifugation time is 10 minutes, take it out and cool it at -20°C until the water phase and the wax layer organic phase are separated, so that the upper layer of the centrifuge tube The wax sheet is fully hardened, tilt the centrifuge tube, and use a sterilized syringe to suck out the tissue mixture solution containing DNA at the bottom to obtain sample A;

[...

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Abstract

The invention discloses a method for extracting DNA from breast cancer paraffin-embedded tissue. The method comprises three steps of deparaffinating, digesting and extracting and precipitating. According to the extracting method, DNA with enough purity and quantity can be obtained with fewer steps, and the method can be effectively used for PCR amplification and DNA sequencing. The method is easy to execute and suitable for large scale production. The prepared DNA bodies are stable in property and high in purity and have very high application value for breast cancer diagnosis. A new concept is provided for preparing of DNA in the breast cancer paraffin-embedded tissue.

Description

technical field [0001] The invention relates to a method for extracting DNA, in particular to a method for extracting DNA from paraffin-embedded tissues of breast cancer. Background technique [0002] Formalin-fixed paraffin-embedded tissues, as the most convenient source of clinical materials, are often used in retrospective studies of large cases. In recent years, with the emergence of many molecular markers related to disease diagnosis, prognosis and treatment, molecular-level gene detection and screening using paraffin tissue sections as materials has become a routine method for clinical diagnosis and related research. For various genetic analysis methods, it is important to first obtain sufficient purity and quantity of DNA. The traditional phenol / chloroform extraction method is the earliest method to purify DNA from paraffin-embedded tissues of breast cancer. Although it can meet the analysis requirements such as PCR, the reagents used are highly toxic, the operation ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 应国光刘博
Owner TIANJIN YINGSHIBO TECH DEV
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