HI, I am using the TRIzol reagent for RNA extraction from yeast. I am planning to shift to a kit based protocol for better reproducibility and consistency. I looked for many RNA extraction kits and Qiagene's RNeasy, Fermentas Genejet and Life technologies Ribopure seem to be good options. The first two use silica column and the work flow is relatively straight forward without the need of any additional equipments. The entire work flow is at room temperature. The Ribopure work work flow is relatively more tedious based on solvent extraction and subsequent glass-fiber filter based purification. Cold temperature is required at initial steps and the work flow is more tedious. Has anyone experienced both the technologies and can tell me which one is better to go for?
Thanks
Hi Syed,
our innuPREP RNA Mini Kit works out fine.
FYI: we use a new chemistry (DC Technology) on traditional spin columns.
The kit eliminates the need for using highly toxic ß-mercaptoethanol; genomic DNA is removed via prefiltration step rather than DNAse treatment.
see http://www.analytik-jena.de/en/life-science/products/prod/cat/isolation-of-total-rna/prod/innuprep-rna-mini-kit.html
For high quality and good recovery Qiagen kits are best, In our lab we use them. E.Z.N.A. kit is good value for money though I did not got good recovery from them sometimes( I was using free sample kits).
Hi, Thank you all for the inputs and suggestions. I have almost standardized the workflow with TRIzol now and am getting pretty good quality RNA now (A260/280~2 with decent bands) and cDNAs are also giving good Cts and no-RT shows no bands. So, thanks again. As of now I will stick to the TRIzol but your inputs will definitely help in future.
Thanks
Our lab has been using the zymo "Direct-zol" RNA kit with great success. You still lyse your cells in trizol, but can apply the trizol directly to the column. We get great yields, purity, and reproducibility.
Can anyone recommend a kit for extraction of total yeast RNA that does not require mechanical disruption?
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