I am not working on  same cells system. But according to my opinion, you can try Trizol Reagents [ Ambion company ] for RNA isolation. Even you can store your cells in trizol rather than in lysis buffer .

We use trizol and isolate 2-4 ug of total RNA (in 20 ul of water) from 1 million of CD14+ monocytes. 260:280 ratio is between 1.8 and 2.0. No problems with storage in trizol as well (-80C). We convert RNA to cDNA immediately after RNA isolation. In the past I was using Quagen RNeasy kit and also didn't observe any problems.

Hi Nikhil.

1 million cells is quite a decent amount. I highly recommend using a RNeasy minikit (Qiagen cat no. 74104). I routinely use this kit to isolate RNA from sorted cells. I commonly sort around 100,000 cells and using this kit recover approx 50ng/ul in 12ul (600ng). Using this kit you should be able to significantly increase your yield. The RNA quality is also extremely good so it allows your downstream applications to be extremely efficient. I would steer clear of TRIzol if you haven't used it before as there is a chance that you might loss your tiny pellet of RNA while doing washes, especially if your pellet is clear.

Thanks Jon......I got the same advice from a lot of people to use this kit specifically....I am going to do the same :)

Hi Nikhil,

if you want to improve the quality and quantity of your RNA consider also the opportunity to use RNeasy MICROKit from Quiagen.  (https://www.qiagen.com/be/products/catalog/sample-technologies/rna-sample-technologies/total-rna/rneasy-micro-kit/)

Hi, Nikhil Kulkarni. We used  trizol and always isolate 1-2 ug of total RNA (in 10 ul of water) from 1 million of CD14+ monocytes. We usually store our cells in trizol and we used  isopropanol precipitation  methods instead of any kit. You can try it.