摘要
以8个烤烟栽培品种为试验材料,对烟草基因组DNA的提取方法进行比较,并对建立烟草SRAP-PCR反应体系的影响因子设置梯度试验.结果显示:改进的CTAB法能较好地满足烟草基因组DNA的提取,在20μLSRAP-PCR的反应体系中,DNA模板40ng、Mg2+1.5mmol/L、dNTP0.25mmol/L、引物0.2μmol/L、Taq聚合酶1.5U,8个烤烟品种的扩增多态性高,带型清晰,差异明显.
Method of DNA extraction and factors influencing SRAP analysis were studied using eight tobacco cultivars. The results showed that, improved method of CTAB contributed to a better extraction of genomic DNA of tobacco. A reliable PCR reaction system for detecting SRAP was developed, each 20μL PCR reaction mixture consisted of 40 ng of genomic DNA, 0.25 mmol/L ofdNTPs, 1.5 mmol/L of Mg^2＋, 0.2 μmol/L of primer and 1.5 unit ofTaq polymerase, every amplification out of the eight cultivars was effective, clear and their differences were significant.
出处
《湖南农业大学学报（自然科学版）》
CAS
CSCD
北大核心
2009年第3期257-259,共3页
Journal of Hunan Agricultural University(Natural Sciences)
基金
云南省大理自治州烟草公司项目(20060678)
关键词
烟草
DNA提取
SRAP
优化
tobacco
DNA extraction
sequence related amplified polymorphism
optimization
