山核桃花芽总RNA提取方法研究

以山核桃的花芽为材料,利用改良CTAB法、异硫氰酸胍法和改良热酚法提取其总RNA,并对RNA产率、纯度、电泳图谱和RT-PCR进行分析,结果表明:改良CTAB法明显优于其它两种提取方法,用它提取的28S rRNA、18S rRNA条带清晰、稳定,无降解,OD260/OD280值保持在1.9～2.0,产率相对较高;经RT-PCR检测发现,改良CTAB提取的RNA能被成花基因特异引物扩增出清晰的条带,说明该法完全适合于山核桃花芽总RNA的提取,并可用于Northern杂交、cDNA文库构建等分子生物学操作.

Methods for RNA Isolation from Flower Buds of Carya cathayensis

Experiment was carded out on isolation of total RNA from flower-bud of Carya cathayensis by the methods of modified CRAB, guanidine isothiocyanate and modified hot phenol. Analysis on isolated RNA yield, purity, electrophoresis profiles and RT-PCR detection demonstrated that CTAB was a better method for the isolation. The result showed the ratio of OD260/OD280 of total RNA isolated by modified CTAB was 1.9 ～ 2.0 with higher yield. The experiment resulted that total RNA obtained by modified CTAB had 2 clear bands 28S rRNA and IgS rRNA without decomposition. RT-PCR showed that one clear band was amplified by using floral gene as special primers from the RNA isolated by the method of CTAB, indicating that method of CTAB was the best for total RNA isolation of C.cathayen~is flower bud, and be used for Northern blotting, cDNA library construction.