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...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.展开更多
文摘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.
