I hope you are adding RNase inhibitor and working on ice or cold temperature. 

What is your method of RNA purification? Sometimes addin a small amount of polyacryl carrier can improve RNA yield dramatically. Have a look here http://www.mrcgene.com/polyacry.htm.

Also In case you store your cells you have  to either do it in RNALater reagent or store quicky at -80oC.

Jivko and Prabodh, I purified RNA with Trizol. I use to do this a lot with other type of cells, but with neutrophils I have had problems. I heard about glycogen to improve RNA yield, but I'll test polyacryl carrier. In relation to use RNAase inhibitor I haven't, but I always work on ice and store my samples in -80oC, althoug I use to perform my cDNA in the same day.

Thanks

you can try to add beta-mercaptoetanol up to 2% to the extraction buffer.

Neutrophils are smaller than many other cell types and have an inherently lower quantity of RNA.  They also are chock full of "ases".  You need to increase the number of cells as your starting point and make sure you add RNAse inhibitor.  You definitely need to work on ice and store all in -80, so that is a good starting point.  I have also syringed them rather than rely on the Trizol or RNA Stat to lyse them.  Hope this helps!

In order to extract RNA from Neutrophils, I work with RNA extraction kits which are more sentitive than the usual ones. I normally choose the RNAqueous Microkit from Ambion to work with them and it works quite well! Anyway, if I use the High Pure RNA Isolation Kit from Roche (this is the one I normally use for other cell subsets) what I have to do is to increase the number of neutrophils I start from (2 millions is normally enough). I have never used the Trizol reagent, but I would try to increase the starting amount of neutrophils or changing the reagent. With the Ambion kit, I have even started from 250.000 neutrophils and it still works!

Mary A Lokuta and Mary A Lokuta  thank you so much. I will try to use some extraction kit. I think in my lab we have RNase from Qiagen....Thank you again.

Hello, did you solve your problem? I'm having the same problem. I'm working with bovine neutrophils 1x10^6 cells and I just get around 4 ng/ul. I'm using TRIzol and RNAeasy.

Could you share your protocol? Thanks

Hi Susan Flores, I hope this answer is not too late. We extract RNA from mouse and human PMNs. We start with 1.5 x 10 ^6 and get around 60-70 ng/uL of RNA concentration using Qiagen RNAeasy Kit.