Protocol: A simple phenol | 合法醫療器材資訊網

由MSBox著作·2011·被引用99次—Dissolvethenucleicacidpellet(mostlyRNA)with50-100μLofnuclease-freewater,applyaPCRplatesealandgentlymixthecontentsusinga ...

In deciding to develop a high-throughput RNA extraction method appropriate for the isolation of RNA from hundreds of Arabidopsis samples, over a broad range of developmental stages, we first examined the suitability of TRIZOL, as it had been described in previous high-throughput nucleic acid extractions [2] where it was favoured for its ability to simultaneously extract both DNA and RNA in a small number of steps. Using TRIZOL we could always isolate high yields of total RNA from Arabidopsis, but found unusual mRNA expression patterns in northern blots prepared using RNA from plants at different stages of growth, specifically low or undetectable expression of several mRNAs in younger seedling stages.

To explore this observation we extracted RNA from Arabidopsis seedlings grown at ambient temperature for three, six or twelve days followed by six weeks of cold treatment before immediately harvesting en...