But seriously, Trizol. Follow your standard trizol extraction protocol, ~100-200 mg tissue to 1ml trizol, homgenize as desired. And workup as typical.

What is your end target? Virological tests?

Hello

You can following this protocol :

1. Homogenize tissue thoroughly and quickly in 1 ml Trizol.

*For insect samples with exoskeletons, a micro-pestle in a 1.5ml microcentrifuge tube works well. It is usually easier to homogenize the tissue in 500ul Trizol, and then add the remaining 500ul to wash the pestle at the end.

*wipe down pestles and leave to evaporate in a hood for at least 24 hours before removing and washing.

2. To increase yield, leave to sit overnight at room temperature in a hood. This step is optional and depends on your application.

3. Phase separation: add 200ml cold chloroform and cap the tube securely. To get the full volume, coat the pipet tip by carefully pipetting the chloroform up and down once first.Shake the tube vigorously by hand for a full 15 seconds. Let sit at room temperature 2-3minutes. You should see the two mixtures separating almost immediately: pink on the bottom and clear on top

4. Centrifuge the mixture 15 min centrifugation at 12,000 x g and 4˚C. Any denatured proteins might appear as a white interface. Avoiding this interface, carefully move the clear top liquid to a new clean, labeled tube. Leave behind some sample if needed.

*** To isolate DNA: discard all of the interface and remaining clear liquid. Close the tube and save the bottom liquid for later DNA isolation. Make sure it is well labeled.

5. Precipitate the RNA by adding 500ul cold isopropanol. Incubate overnight in a 4˚C fridge (Optional but increases yield. Otherwise let sit for 5 minutes- 1 hour at RT or in fridge. Leaving to precipitate in the -80 is not recommended because it might cause contaminants to co-precipitate).

6. Pellet the RNA precipitate by spinning 10 min centrifugation at 12,000 x g and 4˚C. You may see the pellet as a clear, gel –like ball at the base of the tube, or a small white smear. You could see nothing. Discard supernatant by tipping out and blotting the rim. Keep an eye on the pellet (if you can) to make sure it doesn’t get tipped out.

7. Wash the RNA pellet 2 times using 1ml of 75% ethanol (made fresh weekly). Invert the tube (or vortex) each time to rinse all of its surfaces. Between washes, spin the sample at 7,500 x g for 5 minutes at 4˚C.

8. After the final wash, remove as much of the ethanol as possible. Use a pipette if the pellet is visible or carefully tip out the liquid and blot the tube edge. Let air dry until all liquid is gone.Be careful not to over-dry the samples, as they will not go back into solution. If you are walking away to wait for them to dry, cover the open tubes with a kimwipe. Or leave upside down on a clean kimwipe to dry.

9. Re-suspend the pellet in 30-50 ul of Sigma water, or a 0.5%SDS solution for longer storage.

10. Check that all tubes are well labeled with the sample number, date, and your initials. Store at -80˚ C, or in fridge if needed in the next few days.

Ref : http://homepages.eawag.ch/~vorburch/Files/RNA_extraction_TRIZOL_Insects_ABD_April2016.pdf

It depends on your downstream applications. I have routinely used kits (both Picopure and RNAqueous) for extracting RNA for microarray analysis and for RNASeq. Trizol will work well though if you are on a budget.

Some of the column methods do clog if you use too much tissue

Thank you all of you for your generous replies. I will try the mentioned protocol and hope for good results.

Hello, i send you two doc about mosquito grinding process and RNA isolation from mosquito grind liquid, also i would like to notified that the first doc is in French, but easily to understand.

Best regards

Hi. Dr. Prabhakar Mishra

this is good protocol

RNA extraction from Trizol. Protocol for insects

https://homepages.eawag.ch/~vorburch/Files/RNA_extraction_TRIZOL_Insects_ABD_April2016.pdf