Tips and tricks for RNA isolation

Want to become an RNA extraction expert? Read on to discover how to obtain spotless RNA with Zymo Research Corp 's tips and tricks.

RNA extraction is a challenging process with various obstacles, including RNA degradation, low yield, poor purity, and DNA contamination. To address these challenges and maximise the recovery of high-quality, DNA-free RNA, top scientists have shared their best practices and tips.

Stabilising RNA after collection is crucial to prevent degradation. Methods such as immediate solubilisation in an RNase-inactivating lysis buffer or immersion in a stabilisation reagent like DNA/RNA Shield are recommended. These approaches help preserve RNA integrity, particularly for researchers working with field samples or precious patient samples.

Complete sample lysis is essential to maximise RNA yield and quality. Different sample types require specific lysis methods. While detergent-based lysis buffers are commonly used, adding a mechanical lysis step (e.g. bead beating) or an enzymatic lysis step (e.g. proteinase K) can improve lysis efficiency. Complete sample lysis also ensures smooth extraction procedures and avoids issues like column blocking, buffer carryover, DNA contamination, and incomplete elution.

Eliminating DNA contamination is crucial to obtain accurate RNA quantification and reliable downstream analysis. Methods such as TRIzol? phase separation, DNA removal columns, and DNase treatment are recommended. Visualising RNA samples using techniques like agarose gel electrophoresis or qPCR can confirm the absence of high molecular weight DNA fragments.

Choosing the right RNA isolation kit is vital, as one size does not fit all.

Zymo Research offers a range of RNA isolation kits tailored for different sample types and input amounts. These kits employ innovative buffer and column systems to efficiently extract RNA while minimising DNA contamination. They also include on-column DNase treatment, eliminating the need for post-extraction clean-up steps.

Mastering RNA extraction involves three key tips: sample stabilisation, thorough sample lysis, and DNA elimination. By following these tips and using appropriate RNA isolation kits, you can obtain high-quality RNA suitable for any downstream application.

Want to learn more? For more valuable insights and practical recommendations for achieving optimal RNA extraction results, read the full article on Zymo's website here.