Hi Andra Dingman , I am working with a tiny tissue that has about 50 cells and I agree it is a challenge to extract enough RNA from those tissues. What I often do is collect about 50-70 tissues (so 2500-3500 cells) in trizol, then either flash freeze in liquid nitrogen or proceed immediately to RNA extraction. For extraction step, I added trizol to a total volume of 1 mL, vortex/homogenize for 15 seconds => add 200-300uL of chloroform => vortex for another 15 seconds => incubate on ice for 5 minutes. Later, centrifuge at 12,000 x g for 10 minutes at 4oC => Transfer the upper aqueous phase to a cold 1.5mL centrifuge tube and mix with a same volume of isopropanol. Later RNA purification steps were done using RNeasy kit from QIAGEN (74106) following manufacturer's protocol. For the elution step, I often add 20uL nuclease-free water and get a concentration of about 20-25 ng/uL. I did use this method to extract RNA for qPCR, RNAseq and could make them work. Of course, as usual, be extra careful when handling any equipment and avoid RNAse contamination. Hope this help :)

In the very first step, add 10 microliter Beta-mercaptoethanol to 1 ml RLT-plus buffer before use. Use the same buffer accordingly with the table 1 in the kit protocol, Thaw the tissue on ice and homogenize in the buffer. hope it will work

I suggest you go for non-column good old technique, like TRI-zol followed by ethanol/isopropanol precipitation of RNA? May work on small amount sample. Good luck