Method for separating deoxynucleotide from salmon semen
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A deoxynucleotide and semen technology, applied in the field of separation of deoxynucleotide, can solve the problems of insufficient efficiency of the separation method, long time required for separation, prone to errors, etc., and achieve strong promotion value, convenient operation, and high purity Effect
Inactive Publication Date: 2021-07-13
瑞吉明(山东)生物科技有限公司
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[0003] Most of the methods for isolating deoxynucleotides from salmon semen on the market are traditional purification, separation, filtration and other methods, which require more steps in the separation process, and the separation takes a long time , the staff need to operate in strict accordance with more separation steps, if not paying attention, it is easy to make mistakes, the separation method is not efficient enough, and there are certain limitations in application
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[0034] The invention provides a technical solution: a method for separating deoxynucleotides from salmon semen, the separation method comprising the following steps:
[0035] S1, prepare fresh or frozen salmon semen tissues, place them in a glass homogenizer, add a certain amount of cell lysis buffer, and homogenize until no tissue clots are seen, then transfer to a centrifuge tube and add proteinase K to make it Mix well, bathe in a constant temperature water bath for a fixed time, and intermittently oscillate the centrifuge tube several times to initially obtain a mixed liquid;
[0036] S2, the mixed liquid is placed in a desktop centrifuge for centrifugation, and the clear liquid can be obtained and placed in a new centrifuge tube;
[0037] S3, add twice the amount of isopropanol, stir with the clear liquid, and you can see obvious filaments, then use the tip to suck it up, wait for it to dry naturally, and then redissolve it with TE buffer;
[0038] S4, adding an equal am...
example 2
[0048] The invention provides a technical solution: a method for separating deoxynucleotides from salmon semen, the separation method comprising the following steps:
[0049] S1, prepare fresh or frozen salmon semen tissues, place them in a glass homogenizer, add a certain amount of cell lysis buffer, and homogenize until no tissue clots are seen, then transfer to a centrifuge tube and add proteinase K to make it Mix well, bathe in a constant temperature water bath for a fixed time, and intermittently oscillate the centrifuge tube several times to initially obtain a mixed liquid;
[0050]S2, the mixed liquid is placed in a desktop centrifuge for centrifugation, and the clear liquid can be obtained and placed in a new centrifuge tube;
[0051] S3, add twice the amount of isopropanol, stir with the clear liquid, and you can see obvious filaments, then use the tip to suck it up, wait for it to dry naturally, and then redissolve it with TE buffer;
[0052] S4, adding an equal amo...
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Abstract
The invention discloses a method for separating deoxynucleotide from salmon seminal fluid, which comprises the following separation steps: S1, preparing fresh or frozen salmon seminal fluid tissue bodies, putting the tissue bodies into a glass homogenizer, adding a quantitative cell lysis buffer solution, homogenizing until no tissue clots exist, then transferring the tissue bodies into a centrifugal tube, adding protease K, uniformly mixing the tissue bodies and the protease K, and carrying out centrifugal separation; and S2, performing water bath in a constant-temperature water bath kettle for a fixed time, and intermittently oscillating the centrifugal tube for a plurality of times to preliminarily obtain mixed liquid. According to the method, the deoxynucleotide component in the salmon semen can be separated through the steps of oscillation, separation and the like by adopting a dissolution and centrifugal separation purification mode and adding quantitative protease K, a TE buffer solution, absolute ethyl alcohol and the like, so that the deoxynucleotide separated by the separation method is relatively high in purity and relatively high in purity. Compared with the traditional separation method, the method is simpler and clearer, is very convenient for personnel to operate, can efficiently and simply complete the separation work of deoxynucleotide, and has higher popularization value.
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to a method for isolating deoxynucleotides from salmon semen. Background technique [0002] Salmon belongs to the deep-sea fish salmon, which is a kind of deep-sea fish and a very famous anadromous fish. It lays eggs in the streams and rivers in the upper reaches of freshwater rivers, and then returns to the sea for fattening. Also known as deoxyribonucleic acid, it is a small molecular compound composed of purine or pyrimidine bases, deoxyribose and phosphoric acid. It can be isolated and extracted from salmon semen. [0003] Most of the methods for isolating deoxynucleotides from salmon semen on the market are traditional purification, separation, filtration and other methods, which require more steps in the separation process, and the separation takes a long time , The staff need to strictly follow more separation steps to operate, and mistakes are prone to occur if they are not pay...
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