Method for rapidly identifying ploidy of wax gourds by using flow cytometry
The invention provides a method for rapidly identifying the ploidy of wax gourds by using flow cytometry, which comprises the following steps: A, collecting tender leaves, washing and sterilizing the tender leaves, adding pre-cooled and diluted improved lysate, and cutting up the tender leaves by us...
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Main Authors | , , , , , |
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Format | Patent |
Language | English |
Published |
02.09.2015
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Subjects | |
Online Access | Get full text |
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Summary: | The invention provides a method for rapidly identifying the ploidy of wax gourds by using flow cytometry, which comprises the following steps: A, collecting tender leaves, washing and sterilizing the tender leaves, adding pre-cooled and diluted improved lysate, and cutting up the tender leaves by using a pre-sterilized sharp cutter; B, filtering the cut wax gourd leaves which are soaked into the lysate in advance into a centrifugal tube, putting filtrate in a centrifuge, carrying out centrifugalization at room temperature, pouring away supernatant, adding pre-cooled lysate, uniformly stirring, adding dye liquor, uniformly mixing, and carrying out lucifugal dyeing, so that a sample is obtained; and C, through the adoption of the stability of a flow cytometer, calibrating each fluorescence channel by using fluorescent microspheres, and ensuring that the variation coefficient of a fluorescent signal collected by each fluorescence channel is less than 2%; selecting a diploid wax gourd with known ploidy as a contrast sample; and putting a to-be-detected sample on a machine to detect, and collecting 4000-5000 cell nucleuses, so that a flow scatter diagram of the sample and a fluorescence peak diagram of the sample are obtained. According to the invention, a process from sample preparation to result analysis is implemented only in 40 min. |
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Bibliography: | Application Number: CN20151214784 |