An improved method of isolation of high quality total RNA from purple-fleshed sweetpotato,Ipomoea batatas(L.)Lam.
To isolate high quality total RNA from plants of purple-fleshed sweetpotato(Ipomoea batatas L.Lam)is difficult because these plants contain high levels of carbohydrates and pigment compounds.In this study we developed a quick and reliable method for total RNA extraction.Using this extraction protocol,which was derived from the traditional CTAB method with modification,we obtained high quality total RNA from all the tested tissues including leaf blade,petiole,stem,fibrous root,thick root and storage root.Spectrophotometric analysis,together with agarose gel electrophoresis analysis of extracted total RNA,indicated that the total RNA was perfectly intact and the yields were all more than 0.2mg/g fresh weight of different tissues.A metallothionein-like protein gone(G14)fragment was successfully amplified by reverse-transcription-PCR and cDNAs of two target genes(F3H and DFR)were isolated by the method of 3-RACE and 5-RACE using the first-strand 3-RACE-ready and 5-RACE-ready cDNA as templates which were synthesized from the total RNA extraction,indicating that the total RNA extraction was suitable for further molecular analysis.
RNA Isolation RT-PCR Purple-fleshed Sweetpotato(Ipomoea batatas(L.)Lam.)
Wei Zhou Yifu Gong Qili Feng Feng Gao
Guangdong Key Lab of Biotechnology for Plant Development,School of Life Sciences,South China Normal Guangdong Key Lab of Biotechnology for Plant Development,School of Life Sciences,South China Normal
国际会议
3rd China-Japan-Korea Workshop on Sweetpotato(第三届中日韩甘薯学术研讨会)
北京
英文
365-371
2008-10-12（万方平台首次上网日期，不代表论文的发表时间）