Does anybody have experience in whole transcriptome amplification from cells? My usual study includes RNA extraction from cells (n=30-300), isolated from frozen tumors by laser-capture microdissection, the subsequent reverse transcription, cDNA ligation, and whole transcriptome amplification. But I would like to perform reverse transcription, ligation and further WTA directly from cells? Did anybody have such experience? As to DNA, I perform WGA directly from cells and the results are excellent!
We used the ARCTURUS PicoPure RNA Isolation kit for barley pollen (http://products.invitrogen.com/ivgn/product/KIT0204) and succeeded with very low sample amounts (about 25 pollen), sufficient for subsequent RNA sequencing. Give it a try.
Thank you, Goetz! What about RNeasy Plus Micro Kit? Did you have experience with this kit?
I have used RNeasy Micro KIt to isolate RNA from laser microdissected cells from muscle biopsies samples. The amount of samples was very low but this kit works pretty well! The amount of RNA was enough to perform microarray experiments previoulsy WTA using Nugen technology.
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