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琼脂糖电泳展示mRNA差异方法的探讨 被引量:8

Methods of Agarose Gel Electrophoresis for mRNA Differential Display
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摘要 RT-PCR reactions, with template cDNA from etiolated seedlings of Yuetai maintainer lines (Oryzasativa L. ) and 2, 3 or 4 random primers, were carried out at anneal temperatures 36℃, 38℃, 40℃, 42℃ and45℃. Products were separated on 2% agarose gel. No products were obtained at both 42℃and 45℃. Most of themain bands obtained at 36℃, 38℃ and 40℃ were similar, but there are some differences in the weak bands. Thus,we select 40℃ as the anneal temperature in the following experiments. The difference of mRNAs, which were isolated from anthers at different developmental stages of Yuetai CMS. and maintainer lines, were displayed by agarosegel. Differential bands were retrieved and re-amplified. Then, products were purified on 3% agarose gel. Every purified band showed single band on 5% polyacrylamide gel. RT-PCR reactions, with template cDNA from etiolated seedlings of Yuetai maintainer lines (Oryzasativa L. ) and 2, 3 or 4 random primers, were carried out at anneal temperatures 36℃, 38℃, 40℃, 42℃ and45℃. Products were separated on 2% agarose gel. No products were obtained at both 42℃and 45℃. Most of themain bands obtained at 36℃, 38℃ and 40℃ were similar, but there are some differences in the weak bands. Thus,we select 40℃ as the anneal temperature in the following experiments. The difference of mRNAs, which were isolated from anthers at different developmental stages of Yuetai CMS. and maintainer lines, were displayed by agarosegel. Differential bands were retrieved and re-amplified. Then, products were purified on 3% agarose gel. Every purified band showed single band on 5% polyacrylamide gel.
出处 《中国水稻科学》 CAS CSCD 北大核心 1998年第3期177-180,共4页 Chinese Journal of Rice Science
基金 国家自然科学基金
关键词 雄性不育系 恢复系 RNA 电泳 MRNA PCR cytoplasm sterile line electrophoresis polymerase chain reaction restorer line RNA
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  • 1Liang P,Nucl Acid Res,1993年,21卷,3269页
  • 2Liang P,Science,1992年,257卷,967页

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