ZHOU Fei,LIU Yan,HUANG Xu-tang,et al.Establishment and Optimization of RT-PCR System for FAD2-1 in Oil Sunflower[J].HEILONGJIANG AGRICULTURAL SCIENCES,2016,(11):8-10.[doi:10.11942/j.issn1002-2767.2016.11.0008]
油用向日葵FAD2-1基因RT-PCR体系的建立与优化
- Title:
- Establishment and Optimization of RT-PCR System for FAD2-1 in Oil Sunflower
- 文章编号:
- 11-0008-03
- Keywords:
- oil sunflower; FAD2-1; RT-PCR
- 分类号:
- S565.5
- 文献标志码:
- A
- 摘要:
- 为了建立向日葵FAD2-1基因高效特异的PCR扩增体系，以油用向日葵86-1为材料提取总RNA，并反转录合成cDNA，以cDNA第一条链为模板对FAD2-1基因全长编码序列的PCR体系进行了优化。结果表明：根据已公布的FAD2-1序列设计3对特异性引物，每对引物设计了5个退火温度，筛选出适于该基因PCR扩增的引物及其退火温度，并将PCR扩增条件中循环数缩短为30次，延伸时间缩短为7 min，此改进提高了PCR反应的效率。
- Abstract:
- In order to establish a specific and efficient RT-PCR amplification system of FAD2-1 gene in sunflower.Total RNA was extracted from oil sunflower 86-1 and reverse transcription were carried out.The first strand of cDNA as template,the PCR system of complete encoding sequence of FAD2-1 was optimized.The results showed that three pairs of specific primers were designed according to the FAD2-1 sequence published. Five annealing temperature were designed in each pair of primers. The appropriate primers and annealing temperature were screened out for PCR amplification of the gene. The cycles number was reduced to 30 times, and the extension time was shortened to 7 min in PCR amplification conditions. This improvement improved the efficiency of PCR reaction.
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备注/Memo
收稿日期：2016-09-20