A processing method suitable for flow cytometric analysis of dried plant tissue
A technology of flow cytometry analysis and processing method, which is applied in the direction of individual particle analysis, particle and sedimentation analysis, preparation of test samples, etc. It can solve the problems of difficult separation, affecting the accuracy of flow cytometry analysis, and long storage time. , to achieve the effect of increasing the scope of application
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Embodiment 1
[0044] This embodiment is the ploidy determination of Solidago canadensis, and the components of the cell nucleus extract used in this embodiment are:
[0045] Tris: 12mMmM; Na 2 EDTA: 1mM; spermine hydrochloride: 0.2mM; KCl: 70mM, NaCl: 10mM; β-mercaptoethanol: 10mM; Triton X-100: 0.3% (vol / vol); Adjusted with NaOH solution, the cell nucleus extract is pre-stored in a -20°C refrigerator for later use;
[0046] Put 50 mg of mature leaves of Solidago canadensis (leaves dried on silica gel) into a pre-cooled plastic petri dish, then add 1 mL of pre-cooled cell nucleus extract, and then use a double-sided blade to chop the leaves. The chopping process lasts for 10 minutes. For more than 10 minutes, first filter with a 30-micron nylon membrane to collect the filtrate containing the nucleus, and then perform a secondary chopping process on the remaining leaves, that is, use a double-sided blade to continue chopping the leaves. The chopping process lasts for more than 10 minutes un...
Embodiment 2
[0050] The present embodiment is the ploidy determination of the yellow warbler, and the components of the cell nucleus extract used in the present embodiment are:
[0051] Tris: 14mM; Na 2 EDTA: 1mM; spermine hydrochloride: 0.2mM; KCl: 70mM, NaCl: 15mM; β-mercaptoethanol: 20mM; Triton X-100: 0.5% (vol / vol); Adjusted with NaOH solution, the cell nucleus extract is pre-stored in a -20°C refrigerator for later use;
[0052] Put 50 mg of the mature leaves of the oriole (dried on silica gel) into a pre-cooled plastic petri dish, then add 1 mL of pre-cooled cell nucleus extract, and then use a double-sided blade to chop the leaves. The chopping process lasts for more than 10 minutes , first filter with a 30-micron nylon membrane, collect the filtrate containing the nucleus, and then perform a secondary chopping process on the remaining leaves, that is, use a double-sided blade to continue chopping the leaves. The chopping process lasts for more than 10 minutes until all the leaves...
Embodiment 3
[0056] This embodiment is the ploidy determination of Solidago canadensis, and the components of the cell nucleus extract used in this embodiment are:
[0057] Tris: 18mM; Na 2 EDTA: 3mM; spermine hydrochloride: 0.2mM; KCl: 70mM, NaCl: 25mM; β-mercaptoethanol: 20mM; Triton X-100: 0.2% (vol / vol); Adjusted with NaOH solution, the cell nucleus extract is pre-stored in a -20°C refrigerator for later use;
[0058] Put 45 mg of mature leaves of Solidago canadensis (leaves dried on silica gel) into a pre-cooled plastic petri dish, then add 1 mL of pre-cooled cell nucleus extract, and then chop the leaves with a double-sided blade with a thickness of 0.1 mm. The chopping process lasts for more than 10 minutes. First filter with a 30-micron nylon membrane to collect the filtrate containing the nucleus, and then perform a secondary chopping process on the remaining leaves, that is, use a double-sided blade with a thickness of 0.1mm to continue chopping the leaves. The crushing process...
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