摘要
以热研2号柱花草(Stylosanthes guianensis Reyan No.2)叶片为材料,利用紫外分光光度计和凝胶电泳法比较SDS-酚抽提法、CTAB法、Trizol试剂盒和柱式植物RNAout试剂盒法提取柱花草总RNA的质量和纯度。结果表明,改良CTAB法和柱式植物RNAout试剂盒法提取的RNA的OD260nm/OD280nm分别是1.85和1.93,OD260nm/OD230nm均大于2.0。凝胶电泳结果表明,改良CTAB法及柱式植物RNAout试剂盒法均有28SrRNA和18SrRNA两条清晰的条带,且无降解。其他两种方法获得的RNA品质较差,有降解和弥散现象。将改良CTAB法和柱式植物RNAout试剂盒法提取的RNA逆转录成cDNA,cDNA能扩增出一条清晰的β-actin基因片段,进一步证明了改良CTAB法和柱式植物RNAout试剂盒法提取的总RNA具有很高的纯度,其中柱式植物RNAout试剂盒法的效果好于改良CTAB法。
The modified SDS phenol extraction method, Modified CTAB method, common Trizol and Col- umn Plant RNAout were used to extract total RNA in leaf of Stylosanthes guianensis Reyan No. 2, respectively. The quality and quantity of total RNA from above-mentioned methods were compared to select the better methods by UV spectrometer and gel electrophoresis. This study indicated that the value of OD260nm/OD230nm of RNA extracted by modified CTAB method and Column Plant RNAout method were higher than 2.0 and the value of 0D260nm/OD280nm of RNA extracted by modified CTAB method and Column Plant RNAout were 1.85 and 1.93, respectively. Gel electrophoresis showed that RNA extracted by modified CTAB had clearer bands of 28S rRNA andl8S rRNA and they did not degrade, and that RNA extracted by Column Plant RNAout had two clearer bands of 28S rRNA andlSS rRNA and they did not degrade however, RNA extracted by other two methods degraded and dispersed to some degrees. RNA ex- tracted by modified CTAB method and Column Plant RNAout could be reversed to cDNA. The cDNA was amplified and one clear bands of ^-actin gene fragment was observed in agarose gel. These results further demonstrated that the quality and purity of the total RNA extracted by modified CTAB and Column Plant RNAout could be applied into molecular biology experiment, and quality and purity of the total RNA by Column Plant RNAout method were better than that by modified CTAB method.
出处
《草业科学》
CAS
CSCD
北大核心
2011年第7期1326-1330,共5页
Pratacultural Science
基金
广东省科技计划项目(2010B020305011)
