Protocol for ethanol precipitation of nucleic acids

Ethanol precipitation

Generic protocol for ethanol precipitation

🚛 Reagents needed 📦 :

The following reagents are needed. 

Protocol

1️⃣Make ethanol dilutions and store in -20°C (do the same for 80%):

2️⃣Add 2 μl carrier (e.g. linear acrylamide Cat #9520) to the nucleic acid solution and mix well (only needed if you expect a small invisible pellet or low yield).

3️⃣Add 1:10 volume of 3 M sodium acetate and mix thoroughly; for 230 μl of sample, this will be 23 μl of 3 M sodium acetate.

4️⃣Add 4 volumes of 100% ethanol; i.e. 1 mL 100% ethanol for 230 μl sample plus 23 μl 3 M sodium acetate.

5️⃣Mix thoroughly and incubate at –20° C for 1h (long nucleic acids) or overnight (16 hr) for short nucleic acids.

6️⃣Microcentrifuge at top speed for 30 min at 4° C or room temp.

7️⃣Carefully remove and discard the supernatant.

8️⃣Wash the pellet by adding 500 μl 80% cold ethanol. Microcentrifuge at top speed for 10 min at 4°C or room temp, and carefully remove and discard the 80% ethanol.

9️⃣Repeat the 70% ethanol wash if needed to remove the large salt pellet from the sample.

🔟To remove the last traces of ethanol, quickly re-spin the tube, and aspirate any residual fluid with a very fine tipped pipette. Air dry the pellet. Dont let it over dry!
Make 0.1x TE for long-read sequencing buffer storage and elute in this: