Dear fellow researchers,
I am planning to do transcriptomic analyses on population of S. cerevisiae containing very low numbers of individuals (FACSed rare cells, typically # =1000 to 10000 yeast cells), using quantitative Reverse Transcription PCR.
I have seen a lot of different kits for RNA extraction and RT for low numbers of mammalian cells and also one-step kits "Cells-to-Ct" to avoid loosing too much RNA during the extraction process.
However, I haven't found any litterature reporting kits or protocols to achieve this purpose on such a low number of -yeast- cells (which have a lower quantity of RNA compared to higher eukaryotes).
Does anyone have advices for performing qRT-PCR on 103-104 of yeast cells ? Which kit should I use (with or without phenol-chloroform, with RNA extraction/purification or one-step kits, should I go through RNA amplification)?
Thank you for your inputs !
Best,
Théo
pl see the link for answer
http://jme.endocrinology-journals.org/content/29/1/23.short
this link is useful
https://www.ncbi.nlm.nih.gov/pubmed/12200227
regards
Thank you for your inputs, especially Florian!
Indeed I want to quantify the levels or mRNA of different yeast populations of low individual numbers (a bit more than you).
Any particular beads to recommand for the lysis step?
Concerning the kits, I guess I will have to try several differents and see which give the best results on low number of yeast cells.
Thank you again.
Best,
Théo
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