Preparation method of flow cytometry sample for chrysanthemum
A flow cytometry and sample preparation technology, applied in the preparation of test samples, etc., can solve the problems of complex origin and genetic background, and the inability to obtain ideal results, so as to improve accuracy, protect DNA from degradation, and prepare The effect of process reduction
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[0032] A flow cytometry sample preparation method applicable to chrysanthemum, comprising the following steps:
[0033] The first step, lysing step: lysing, filtering, and centrifuging the chrysanthemum tissue sample to obtain chrysanthemum cell nuclei to be stained.
[0034] Specifically, this step specifically includes the following operations:
[0035] A. Get the chrysanthemum leaves, put them into a petri dish (preferably 6 cm in diameter) filled with pre-cooled cell lysis buffer, and cut the chrysanthemum leaves into pieces within 1 min with a sharp blade;
[0036] The types of chrysanthemums are: chamomile, heterochromia chrysanthemum, Asian chrysanthemum, Maohua chrysanthemum, Korean chrysanthemum, Japanese daisy, artemisia japonica, and purple wild chrysanthemum; the chrysanthemum leaves are selected from the new young leaves of the upper part of the unflowered chrysanthemum;
[0037] The mass volume (w / v) ratio of chrysanthemum leaves and cell lysis buffer is 1-10:10...
Embodiment 1
[0071] A flow cytometry sample preparation method applicable to chrysanthemum, comprising the following steps:
[0072] (1), take 50 mg of chrysanthemum leaves, put them into a petri dish dripped with 1 ml of pre-cooled cell lysis buffer, and cut the chrysanthemum leaves into pieces within 1 min with a Gillette double-sided blade;
[0073] The types of chrysanthemums are: chamomile, heterochromia chrysanthemum, Asian chrysanthemum, Maohua chrysanthemum, Korean chrysanthemum, Japanese daisy, artemisia japonica, and purple wild chrysanthemum; the chrysanthemum leaves are selected from the new young leaves of the upper part of the unflowered chrysanthemum;
[0074] The components of the cell lysis buffer are:
[0075] 15mM MOPS, 2mM Na 2 EDTA, 0.5mM spermine.4HCL, 80mM KCl, 20mM NaCl, 0.1% (v / v) TritonX-100, 1% (w / v) pvp-10, 0.1% (v / v) β - mercaptoethanol, pH 7.5;
[0076] (2), then add 1 ml of pre-cooled cell lysis buffer to the fragments to release the nuclei to obtain a nuc...
Embodiment 2
[0106] A flow cytometry sample preparation method applicable to chrysanthemum, comprising the following steps:
[0107] (1), take 50 mg of chrysanthemum leaves, put them into a petri dish dripped with 1 ml of pre-cooled cell lysis buffer, and cut the chrysanthemum leaves into pieces within 1 min with a Gillette double-sided blade;
[0108] The types of chrysanthemums are: chamomile, heterochromia chrysanthemum, Asian chrysanthemum, Maohua chrysanthemum, Korean chrysanthemum, Japanese daisy, artemisia japonica, and purple wild chrysanthemum; the chrysanthemum leaves are selected from the new young leaves of the upper part of the unflowered chrysanthemum;
[0109] The components of the cell lysis buffer are:
[0110] 15mM MOPS, 2mM Na 2 EDTA, 0.5mM spermine.4HCL, 80mM KCl, 20mM NaCl, 1% (v / v) TritonX-100, 0% (w / v) pvp-10, 0.2% (v / v) β - mercaptoethanol, pH 7.5;
[0111] (2), then add 1 ml of pre-cooled cell lysis buffer to the fragments to release the nuclei to obtain a nucle...
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