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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Bibliographic Details
Main Authors LIU CHAOAN, WAN ZHENGLIN, LI LIZHI, ZHOU YANXIA, DENG JIANYING, WU PENG
Format Patent
LanguageEnglish
Published 02.09.2015
Subjects
BEER
BIOCHEMISTRY
CHEMISTRY
COMPOSITIONS OR TEST PAPERS THEREFOR
CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES
ENZYMOLOGY
INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES
MEASURING
MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS
METALLURGY
MICROBIOLOGY
MUTATION OR GENETIC ENGINEERING
PHYSICS
PROCESSES OF PREPARING SUCH COMPOSITIONS
SPIRITS
TESTING
VINEGAR
WINE
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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.
Bibliography:Application Number: CN20151214784