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Nile tilapia sex chromosome DNA (deoxyribonucleic acid) library construction method

A Nile tilapia and DNA library technology, applied in the field of molecular biology, can solve the problems of lack of operation procedures and technical specifications, high requirements for production technology operation, achieve high repeatability, improve production efficiency, and reduce separation difficulty Effect

Inactive Publication Date: 2017-05-31
FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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AI Technical Summary

Benefits of technology

This technology allows for easier understanding on how it works compared with current methods such as cell culture or flow cytometry techniques. It also makes better use of certain types of DNA (chromatin) during different stages of development. Additionally, this process helps identify new genotypes more accurately through analysis of their characteristics at various times throughout its lifecycle.

Problems solved by technology

Technically speaking, it describes various techniques described in the patents relates to identifying and studying tuna (Cichlidae) DNA sequences during mitosis. However, current methods have limitations like being complicated processes requiring expensive apparatusing devices, making them hard to use widely across all species. There also exist challenges associated with obtaining accurate data from these samples obtained via conventional analysis tools.

Method used

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  • Nile tilapia sex chromosome DNA (deoxyribonucleic acid) library construction method
  • Nile tilapia sex chromosome DNA (deoxyribonucleic acid) library construction method
  • Nile tilapia sex chromosome DNA (deoxyribonucleic acid) library construction method

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Embodiment 1

[0039] A method for constructing a Nile tilapia sex chromosome DNA library, comprising the following steps:

[0040] (1) Nile tilapia head and kidney cell culture: Take Nile tilapia with a weight of more than 50 g and drain the tail for 20 minutes, then put the Nile tilapia into a dissecting tray, and use clean scissors to cut open the fish From the back of the head, take out the head kidney with tweezers, put the head kidney tissue into a culture dish filled with 0.8% normal saline, wash it 2-3 times with normal saline, put a 100-mesh cell sieve in a culture dish filled with 10mL normal saline. Put the head kidney tissue on the sieve in the culture dish, grind it gently with the smooth pusher in the syringe, rinse the sieve with a small amount of normal saline, and wash the head kidney cells into the culture dish; then use 150 mesh cells Sieve and filter once, put the cell suspension into a 15mL centrifuge tube, centrifuge at 1200rpm for 10min, discard the supernatant, add 12...

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Abstract

The invention relates to a Nile tilapia sex chromosome DNA (deoxyribonucleic acid) library construction method, belonging to the technical field of molecular biology. The method comprises the following steps: Nile tilapia head-kidney cell culture, cell hypotonic treatment, cell fixation, tabletting, chromosome cutting, separation, enzyme digestion and joint connection, amplification and recovery, and segment sequencing and analysis. The method is standardized, and is simple to operate. The proportion of lymphocytes in the chromosome separation phase prepared from the head-kidney cells is higher than that of hemocytes, thereby ensuring the favorable chromosome separation phase in the same visual field, and obviously enhancing the tabletting efficiency. By using the method, the neutral chromosome can be obviously distinguished from other chromosomes in the high power field. The method has the advantages of high cutting stability and high repetitiveness, lowers the separation difficulty of the Nile tilapia sex chromosome, and enhances the library construction specificity.

Description

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Claims

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Application Information

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Owner FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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