荻MsFT1及MsFT2基因的克隆及序列分析

时间:2024-12-26 15:57:20 来源:作文网 作者:管理员

关键词 荻;光周期;Hd3a;FT

Key words Miscanthus Sacchariflor♀us;photoperiod;Hd3a;FT

1 材料与方法

1.1 试验材料及光周期ผ诱导

2 结果与分析

采用改良Trizol法提取荻叶片的总mRNA用琼脂糖凝胶电泳检测后,条带(图2)清晰完整性较好可以进行下一步。设计好的内参基因ACTIN11引物ACTIN11-F:CAAGCAGTAT GAAGATCAAGGT;ACTIN11-R:GCCTGACTCATCATACTCAC对反转后的cDNA进行PCR检测,均得到134 bp左右清晰的亮带,证明得到的cDNA第1条链结果良好,可进行下一步克 ッ隆的操作。cDNA作为扩增的模板,利用5对引物进行PCR扩增经凝胶电泳检测得到a、b、c、d、e共5条亮带,条带大小与预计大小相符。进行切胶回收纯化,载体连接,转入到DH5ฑa感受态扩增❣培养,再经过PCR检测为阳性菌后,每个PCR产物10个重复,送往海东盛生物有限公司进行测序。

3 讨论

4 参考文献

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