A method for extracting dna from a single egg of copepods

Abstract

The invention relates to a method for extracting DNA from a single egg of a copepod, relating to a DNA extraction technique. Provided is a method for extracting DNA from single eggs of copepods. Including individual eggs produced by different egg diameters and different spawning methods. This method can not only ensure the efficiency of DNA extraction as much as possible, but also the extracted DNA can be directly applied to downstream PCR experiments. Place a single copepod egg in 1×TE buffer solution, add proteinase K to make the final concentration of proteinase K reach 2.5-5.5 mg / mL, and obtain a mixed extract; pass the obtained mixed extract through a heat incubation procedure, The crude extract of single egg DNA is obtained; the crude extract of single egg DNA is precipitated by a nucleic acid co-precipitation agent to obtain DNA precipitation. The precipitate can be dissolved in sterilized water and stored in the refrigerator for later use.

Description

technical field [0001] The invention relates to a DNA extraction technology, especially a novel DNA extraction method that is practical, rapid and effective for single eggs of copepods, including different egg-laying modes, different physiological states and different egg diameters. Background technique [0002] At present, there are mainly two methods for extracting single eggs of zooplankton in the world, one is Montero-Pau et al. Application of an inexpensive and high-throughput genomic DNA extraction method for the molecular ecology of zooplanktonic diapausing eggs .Limnology and Oceanography: Methods, 2008, 6: 218-222) proposed the HotSHOT method, this method is mainly aimed at the extraction of a single egg genome DNA of zooplankton diapause eggs, and its operation steps mainly include the following steps: adding certain A large amount of lysis buffer was used; diapause eggs were broken by physical means and tools; finally, genomic DNA was released through a process of...

AI Technical Summary

Problems solved by technology

It is worth mentioning that there are many problems in the two commonly used methods for extracting DNA from single zooplankton eggs and the commercial kits that have emerged in an endless stream. In summary, there are mainly the following points: 1. The two methods that are currently popular Both use physical means and tools to destroy the structure of the egg. This operation will make the amount of DNA extracted less than the theory. These are man-made losses caused by the operation, and the DNA content is not high. For a single egg, this loss is huge; 2. In the method of Chelex-100, the resin added by itself virtually increases the chance of contamination, and practice has proved that the addition of resin has a great impact on downstream PCR experiments. Inhibition; 3. The commercial kit mainly completes the collection and purification of DNA through the process of membrane adsorption and elution. While the quality of DNA is improved, the elution efficiency directly affects the concentration of DNA, which will also directly affect the downstream. PCR experiment

[0003] The above are the research progress in this field abroad, so far, there is no domestic report on the extraction of genome DNA from a single egg of copepods

Images