A Flow Cytometry Identification Method for the Progeny of Homozygous Lines Obtained from Wheat Haploid Breeding
An identification method and wheat technology, applied in the biological field, can solve the problems of low effective cell number, influence detection accuracy, cell activity damage, etc., achieve the effects of good accuracy and reproducibility, improve dispersion efficiency, and prevent detection interference
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Embodiment 1
[0053] A flow cytometric identification method for the homozygous progeny of wheat haploid breeding adopted in this embodiment comprises the following steps:
[0054] S1. Add a certain amount of citric acid and sodium chloride into double-distilled water and mix evenly, so that the concentrations of citric acid and sodium chloride are 420g / L and 180g / L respectively, to obtain a buffer mother solution, and store it at 4°C for later use;
[0055] S2. Mix the double distilled water and the buffer solution mother solution in a volume ratio of 19:1 to obtain the buffer solution for subsequent use;
[0056] S3, adding 0.06ml of nonylphenol polyoxyethylene ether and 0.06ml of β-mercaptoethanol to each 100ml of the buffer solution, and mixing to obtain a buffer working solution for later use;
[0057] S4. Take a certain amount of Hoechst 33258 and add it to the buffer solution to dissolve and mix evenly, so that the concentration of Hoechst 33258 is 200mg / L to obtain a dyeing mother s...
Embodiment 2
[0063] The flow identification method for the homozygous progeny of a kind of wheat haploid breeding adopted in this embodiment is basically the same as that of Example 1, the difference lies in the following aspects:
[0064] Step S3 is: adding 0.1 ml of nonylphenol polyoxyethylene ether and 0.09 ml of β-mercaptoethanol to every 100 ml of the buffer solution, and mixing to obtain a working buffer solution for later use.
Embodiment 3
[0066] The flow identification method for the homozygous progeny of a kind of wheat haploid breeding adopted in this embodiment is basically the same as that of Example 1, the difference lies in the following aspects:
[0067] Step S3 is: adding 0.01ml of nonylphenol polyoxyethylene ether and 0.01ml of β-mercaptoethanol to every 100ml of the buffer solution, and mixing to obtain a working buffer solution for later use.
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