8.4 RNA Isolation Protocol

Unlike DNA isolation, RNA isolation can be time consuming, delicate, and fruitless work at times. While RNA isolation can be a challenging technique to master, it is also a very rewarding skill to have. Before jumping into RNA isolation review the Chapter 4: Sample Collection. Knowing how to handle collected tissue and downstream products is an important means to getting high quality RNA yields that accurately reflect in vivo RNA levels.

8.4.1 RNase Free

RNase is an enzyme that degrades RNA strands. These molecules can be found in the body, on your skin, on the lab bench, and in the air. RNase is also very hard to neutralize (RNase A variant), especially when compared to DNase. Because of this it is essential to use RNase free techniques when preforming any task in the RNA isolation procedure. Many vendors supply DNase, RNase, Nuclease, and Protease free tips, water, tubes, etc. which are also sterile. Other techniques that are helpful include keeping a designated area for RNA work and additional cleaning with RNase Away, an RNase decontaminant. Many of these techniques can be found in this post on Thermo-Fisher’s website.

8.4.2 Kits

• Trizol
• Oligotex
• Quantitect Reverse Transcription (cDNA)