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Kit and method for extracting DNA from micro samples

A kit and sample technology, applied in the field of molecular biology, can solve the problems of low amount of extracted DNA, affecting the speed of analysis and detection, and time-consuming, etc., and achieve the effect of high DNA content, fast extraction speed and high purity

Active Publication Date: 2011-01-05
中生方政生物技术股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the spin column method can extract DNA and ensure its purity, the amount of DNA extracted is generally not high, which cannot meet the requirements of forensic testing, archaeological analysis, and medical testing.
In addition, the spin column method uses some toxic reagents, such as phenol, chloroform, etc., which is not conducive to the health of the experimenters; at the same time, the spin column method consumes a lot of time in the entire process of extracting DNA, which affects the speed of the next step of analysis and detection

Method used

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  • Kit and method for extracting DNA from micro samples
  • Kit and method for extracting DNA from micro samples
  • Kit and method for extracting DNA from micro samples

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Embodiment 1: kit of the present invention

[0079] The kit for extracting DNA from trace samples of the present invention comprises:

[0080] Pretreatment buffer: pH 7.2, containing 135mM NaCl, 2.7mM KCl, 1.5mM KH 2 PO 4 , 8mM K 2 HPO 4 . Specific preparation method: weigh 0.8gNaCl, 0.02g KCl, 0.024g KH respectively 2 PO 4 , 0.18g K 2 HPO 4 , add an appropriate amount of distilled water to dissolve and mix, then adjust the pH to 7.2 with HCl, and then adjust the volume to 100ml.

[0081] Lysis solution: 0.1M NaCl, pH 8.0 10mM Tris-HCl, 25mM EDTA and 0.5% SDS. Specific preparation method: Measure 1ml of 1M Tris-HCl (pH 8.0), 5ml of 0.5M EDTA (pH 8.0), 2ml of 5M NaCl, and 5ml of 10% SDS, add appropriate amount of distilled water and adjust the volume to 100ml.

[0082] Binding solution: 4.5M guanidine isothiocyanate, pH4.4 10mM Tris-HCl, 20% Ttiton-X 100, 10mM Urea, 10mM EDTA and 5% Tween20. Specific preparation method: Weigh 53.172g guanidine isothiocyanate a...

Embodiment 2

[0089] Example 2: DNA Extraction from Dried Blood Spots

[0090] DNA was extracted from dried blood spots using the kit described in Example 1.

[0091] 1. Extraction reagent

[0092] Pretreatment buffer: pH 7.2, containing 135mM NaCl, 2.7mM KCl, 1.5mM KH 2 PO 4 , 8mM K 2 HPO 4 .

[0093] Lysis solution: 0.1M NaCl, pH 8.0 10mM Tris-HCl, 25mM EDTA and 0.5% SDS.

[0094] Binding solution: 4.5M guanidine isothiocyanate, pH4.4 10mM Tris-HCl, 20% Ttiton-X 100, 10mM Urea, 10mM EDTA and 5% Tween20.

[0095] Conditioning binding solution: analytically pure isopropanol.

[0096] Biomagnetic beads: the core is Fe 3 o 4 , Peripheral wrapped silicon dioxide, product of Promega company.

[0097] Magnetic Separation Rack: Built-in magnets.

[0098] Protein removal solution: 6M guanidine hydrochloride, pH7.525mM Tris-HCl and 37% absolute ethanol.

[0099] Rinse solution: pH 7.5 10 mM Tris-HCl, 100 mM NaCl, 25% absolute ethanol and 25% isopropanol.

[0100] Eluent: 10 mM Tris-HCl...

Embodiment 3

[0116] Example 3: DNA extraction from saliva

[0117] 1. The kit for extracting DNA from trace samples according to the present invention

[0118] The kit for extracting DNA from trace samples of the present invention comprises:

[0119] Pretreatment buffer: pH 7.2, containing 135mM NaCl, 2.7mM KCl, 1.5mM KH 2 PO 4 , 8mM K 2 HPO 4 .

[0120]Lysis solution: 0.05M NaCl, pH 6.5 5mM Tris-HCl, 10mM EDTA and 0.25% SDS.

[0121] Binding solution: 3M guanidine isothiocyanate, pH3.5 5mM Tris-HCl, 5% Ttiton-X100, 5mM Urea, 5mM EDTA and 5% Tween20.

[0122] Conditioning binding solution: analytically pure isopropanol.

[0123] Biomagnetic beads: the core is Fe 3 o 4 , Peripheral wrapped silicon dioxide, product of Promega company.

[0124] Magnetic Separation Rack: Built-in magnets.

[0125] Protein removal solution: 2M guanidine hydrochloride, pH6.5 5mM Tris-HCl and 5% absolute ethanol.

[0126] Rinse solution: pH 6.55mM Tris-HCl, 50mM NaCl, 10% absolute ethanol and 10% isop...

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Abstract

The invention relates to the field of molecular biology and discloses a kit and a method for extracting DNA from micro samples. The kit comprises a binding liquid, a regulated binding liquid and a deproteinized liquid, wherein the binding liquid comprises 3-8M of guanidinium isothiocyanate, 5-50mM of Tris-HCl with a pH value of 3.5-5.5, 5-25% of Ttiton-X 100, 5-20mM of Urea, 5-20mM of EDTA and 5% of Tween20, the regulated binding liquid is isopropanol, and the deproteinized liquid comprises 2-8M of guanidine hydrochloride, 5-50mM of Tris-HCl with a pH value of 6.5-8.0 and 5-50% of absolute ethyl alcohol. The DNA extracted by the kit and the method has high content and purity, the extraction speed is high, and no toxic reagent is used. The kit and the method are applicable for extracting DNA from various micro samples.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a kit for extracting DNA from trace samples, and also to a method for extracting DNA from trace samples. Background technique [0002] DNA, translated into Chinese as deoxyribonucleic acid, is the main component of chromosomes. DNA is the most important biological information molecule, which can form genetic instructions to guide the development of organisms and the operation of life functions. Transcribing genes and expressing proteins in an orderly manner in time and space completes all programs of directional development; preliminarily determines the unique traits and personalities of organisms and all stress responses when interacting with the environment. Because DNA plays such an important role in the growth and development of organisms, it becomes an important research object for molecular biology researchers, so the separation and purification of genomic DNA has become o...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 李艳萍
Owner 中生方政生物技术股份有限公司
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