| 摘要: |
| 【目的】探讨榛子总RNA提取的有效方法，为今后开展榛子的分子生物学等研究奠定基础。【方法】以榛子的成龄叶片、幼嫩叶片和雄花芽为试材，采用植物RNA提取试剂盒、Trizol法、改良的CTAB法Ⅰ、改良的CTAB法Ⅱ、CTAB法和改良的SDS法提取其总RNA，通过琼脂糖凝胶电泳、紫外分光光度计和PCR检测所提取总RNA样品的品质。【结果】除了Trizol法，其他5种方法都能提取到总RNA，但是改良的CTAB法Ⅰ更适合榛子幼嫩叶片、成龄叶片和雄花芽总RNA的提取，此方法提得的总RNA较完整，条带清晰，品质较优，28S rRNA亮度约是18S rRNA的2倍，且无降解现象。经RT-PCR验证，以改良的CTAB法Ⅰ提取的RNA为模板扩增得到的条带清晰，与目的片段大小一致，能够满足后续试验的要求。【结论】改良的CTAB法Ⅰ是榛子幼嫩叶片、成龄叶片和雄花芽等组织总RNA提取的最有效方法。 |
| 关键词: 榛子 总RNA CTAB SDS |
| DOI： |
| 分类号: |
| 基金项目:北京林业大学博士后特别资助基金项目（200902052）；国家林业局重点项目（2011-03） |
|
| Comparison of extraction methods for hazelnut total RNA |
|
|
| Abstract: |
| 【Objective】This study aimed to investigate effective methods for hazel total RNA extraction.【Method】Total RNAs were isolated from adult leaf,juvenile leaf and male flower bud of hazelnut through six methods including Plant RNA Kit,Trizol method,modified CTAB methodⅠ,modified CTAB methodⅡ,CTAB method,and modified SDS method.The quality of the total RNA was tested by gel electrophoresis,Ultraviolet spectrophotometer and PCR.【Result】Total RNAs could be obtained from the three tissues by five methods except Trizol method,and modified CTAB methodⅠwas the best method in terms of RNA quality.Based on gel electrophoresis,total RNA extracted by modified CTAB methodⅠhad two clear bands of 28S rRNA and 18S rRNA without degradation,and with relative integrity,high purity and good integrity.The OD260/OD280 ratios of the total RNA were between 1.8 and 2.1.The quality of the RNA was determined by RT-PCR.The PCR product had a distinct band with the same size as theoretical fragment and could satisfy the requirement of following molecular biology study.【Conclusion】The modified CTAB methodⅠwas the best for extraction of total RNA from hazel adult leaf,juvenile leaf and male flower bud. |
| Key words: hazelnut total RNA CTAB SDS |
