Method for quickly extracting genome DNA from fish blood
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A genome and blood technology, applied in the field of genomic DNA extraction, can solve the problems of short storage time, low purity of genomic DNA, and high cost, and achieve the effect of ensuring purity, simplifying the process of extracting genomic DNA, and low price
Inactive Publication Date: 2014-04-30
HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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[0005] The purpose of the present invention is to solve the existing problems of cumbersome extraction process, high cost, low purity and short storage time in the existing process of extracting genomic DNA from fish blood, and to provide a method suitable for extracting genomic DNA from fish blood. A method for rapid extraction of genomic DNA from
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specific Embodiment approach 1
[0028] Embodiment 1: In this embodiment, a method suitable for rapidly extracting genomic DNA from fish blood is carried out according to the following steps:
[0029] 1. Fish blood sample preparation:
[0030] ①. Use a disposable sterile syringe to extract 0.1mL-0.2mL of blood from the tail of the fish;
[0031] ②. Add 20 μL to 30 μL of the blood obtained in step ① into a centrifuge tube containing 800 μL of lysate, and repeatedly invert until the mixture is uniform to obtain a fish blood sample;
[0032] Wherein, the molar concentration of EDTA in the lysate is 90mmol / L~110mmol / L, the molar concentration of Protainase K in the lysate is 100μg / mL~200μg / mL, the sodium lauryl sarcosine in the lysate The mass concentration is 0.5%, and the pH of the lysate is 8.0;
[0033] 2. Place the centrifuge tube containing the fish blood sample obtained in step 1 in a water bath with a temperature of 55°C for 1h to 1.5h for digestion, and then extract it twice with a phenol-chloroform-is...
specific Embodiment approach 2
[0036] Specific embodiment two: the difference between this embodiment and specific embodiment one is that the species of fish described in step one 1. are carp, crucian carp, mandarin fish, grass carp, silver carp, rainbow trout, golden trout, white spot salmon , Shannu trout, yellow catfish, German mirror carp, Qinghai Lake naked carp or Ussuri catfish. Other parameters are the same as in the first embodiment.
specific Embodiment approach 3
[0037]Embodiment 3: The difference between this embodiment and Embodiment 1 or 2 is that the disposable sterile syringe described in step ① is a 0.5mL, 1mL or 2mL disposable sterile syringe. Other parameters are the same as those in Embodiment 1 or Embodiment 2.
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Abstract
The invention relates to a method for quickly extracting a genome DNA from fish blood, relates to a method for extracting the genome DNA and aims at solving the problems in the process of extracting the genome DNA from the fish blood as follows: the extracting process is complex, the cost is high, the purity of the extracted genome DNA is not high, and the preservation time is short. The method comprises the following steps: 1, preparing a fish blood sample: adding blood into a lysis solution for uniformly mixing to obtain the fish blood sample, wherein the molar concentration of EDTA in the lysis solution is 90-110 mmol/L; 2, after the fish blood sample is subjected to digestive treatment, extracting with a phenol-chloroform-isoamyl alcohol mixed liquid; centrifuging to obtain supernate, and then extracting with a chloroform-isoamyl alcohol mixed liquid to obtain blood genome DNA digestive liquid; 3, precipitating the DNA digestive liquid with absolute ethyl alcohol, then washing the precipitate with 70% ethyl alcohol, and drying to obtain a fish blood genome DNA precipitate, namely completing the process of extracting DNA. The method is applied to the technical field of genes.
Description
technical field [0001] The invention relates to a method for extracting genome DNA. Background technique [0002] Blood is an opaque red liquid flowing in the heart and blood vessels. Its main components are plasma and blood cells, which belong to connective tissue. Extract high-quality DNA from blood, and analyze DNA to help researchers better carry out work in the field of biology. The extraction of genomic DNA is a key step in fish genetic engineering technology and related scientific research. Especially for the detection of transgenic fish, there are often significant differences in the quality of DNA from different tissue sources, which is an important factor restricting the subsequent detection of genetically modified fish. We extract DNA from blood, from fin rays, from muscle, from liver, in different ways. [0003] The existing method for extracting genomic DNA from fish blood is to add the collected blood to the lysate, purify it through membrane dialysis, and t...
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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 闫学春栾培贤
Owner HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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