江西农业学报2023，Vol.35Issue(10)：7-13,7.DOI:10.19386/j.cnki.jxnyxb.2023.10.002
大麻叶片基因组DNA快速提取方法与应用
A Rapid Method for Genomic DNA Extraction from Hemp Leaves and Its Application
摘要
Abstract
In order to meet the needs of molecular detection and identification of hemp and marker-assisted screening,gene cloning and sequencing of large-scale breeding materials,CTAB method was used as the control,and rapid extraction methods such as alkali cracking method,TPS buffer method and TE buffer method were compared and optimized.The influencing factors that may affect the yield and quality of DNA extraction were analyzed.The composition and dosage of the extraction solution,the sample size and collection method,the operation and required time of each step in the extraction process,the storage time and the application range were determined,and a microtrace,efficient and rapid extraction method for genomic DNA from hemp leaves was established.Two pieces of hemp leave cut(0.5 cm in diameter)were put into a 2 mL centrifuge tube with 300 μL of TE buffer(10 mmol/L EDTA,150～200 mmol/L Tris-HCl)and 2 steel beads,and then were ground rapidly for 30 s.The ground samples were centrifuged at 13000 r/min for 1 min,then 100～150 μL supernatant was frozen at-18℃for 15 min and melted at room temperature,centrifuged at 13000 r/min for 1 min,and the appropriate amount of supernatant was used as template DNA for PCR amplification.This method is simple,economical,high efficiency,time saving,safe operation and high PCR amplification efficiency,thus suitable to extract samples scalable from one to hundreds,and can be widely used in hemp molecular identification and molecular marker-assisted selection of breeding materials.关键词
大麻/DNA提取/PCR/分子检测Key words
Hemp/DNA extraction/PCR/Molecular detection分类
农业科技引用本文复制引用
张庆滢,李磊,段慧婷,吕品,郭蓉,许艳萍,郭鸿彦,杨明..大麻叶片基因组DNA快速提取方法与应用[J].江西农业学报,2023,35(10):7-13,7.基金项目
国家现代农业产业技术体系(CARS-16-E07) （CARS-16-E07）
云南省农业联合专项重点项目(202301BD070001-146) （202301BD070001-146）
云南省重大科技专项计划子项目(202102AA310041-1). （202102AA310041-1）
