野生玫瑰基因组DNA提取及CDDP引物筛选

摘要

野生玫瑰中多糖多酚含量较高，DNA 提取较为困难，本试验在植物基因组 DNA CTAB 提取法的基础上，通过延长水浴时间、调整氯仿－异戊醇（24∶1）抽提次数为2～3次等一系列方法提高野生玫瑰基因组DNA 的提取纯度。以2×Es Taq MasterMix（含染料）10μL，40 ng／μL DNA 模板1μL，10 pmol／μL 引物1μL， ddH2 O 8μL 为 CDDP －PCR 反应体系，对21条 CDDP 引物进行多态性筛选。结果表明，用改良的 CTAB 法提取的野生玫瑰基因组 DNA 纯度较高；21条 CDDP 引物中有16条对野生玫瑰品种的鉴别能力较高。本研究为野生玫瑰 CDDP 分子标记及后续各种分子生物学研究奠定基础。%DNA extraction of Rosa rugosa is difficult for their high contents of polyphenol and polysac-charide.Here a modified CTAB method for Rosa rugosa DNA extraction was presented through extending water -bath time,appropriately adjusting chloroform isoamyl alcohol (24∶1)extraction time to 2 ~3 times and etc.Twenty -one CDDP polymorphic primers were screened with reaction system containing 10 μL of 2 ×Es Taq MasterMix (including dye),1 μL of 40 ng/μL DNA template,1 μL of 10 pmol /μL primer and 8 μL of ddH2 O.The results showed that the genomic DNA of Rosa rugosa extracted with modified CTAB method owned high purity,and 16 CDDP primers had higher discriminating ability on Rosa rugosa.This research pro-vided bases for CDDP molecular markers and subsequent molecular biology researches of Rosa rugosa.