中文摘要：

目的：建立一个适合于甘蔗（Saccharum officenarum）为研究材料的cDNA-AFLP反应的优化体系。方法：用Rever-tAidTM First Strandc DNA Synthesis Kit反转录获得第一链,用Rnase H、E.coliDNA聚合酶Ⅰ和E.coliDNA连接酶合成双链cDNA,用双低频酶EcoRⅠ和PstⅠ对dscDNA酶切,连接,预扩增,和选择性扩增的关键因素进行分析。结果：500ng的dscDNA双酶切5h,将16℃过夜连接的产物稀释1倍用作预扩增的模板,预扩增产物稀释50倍作为选择性扩增模板,6%聚丙烯酰胺凝胶检测及银染,扩增条带均匀分布,清晰可辨且主要分布在200～2000bp之间。结论：该体系具有稳定性高,重复性好等优点,可用于甘蔗cDNA-AFLP分析。

英文摘要：

Objective：To establish an appropriate cDNA-AFLP analysis system for Saccharum officinarum.Method：With the Revert AidTM First Strand cDNA Synthesis Kit,the first single strand cDNA was synthesized through the reverse transcription from the total RNA.The second strand （ ds） cDNA was synthesized by using RNase H,E.coli DNA polymerase Ⅰ and E.coli DNA ligase.The influencing factors of several reactions,including two low-frequency enzyme EcoRⅠ and PstⅠ digestion,adapter ligation,pre-amplification,selective amplification,were analyzed.Result：500ng cDNA was digested completely by EcoRⅠ and PstⅠ for 5 hours.The clear cDNA-AFLP fingerpritings have been obtained when ligation products were diluted to 1 times for pre-amplification template and pre-amplification products were diluted to 50 times for selective amplification and after 6% polyacrylamide gel electrophoresis and silver staining.The amplication which were centralized between 200 bp and 2000 bp.Conclusion：With the virtue of high stability and good repeatability, cDNA-AFLP system is very fit for Saccharum officinarum.