Isolating high-quality RNA for RNA-Seq from 10-year-old blood samples

Area of Science:

• Molecular Biology
• Biobanking and Sample Preservation

Background:

• RNA analysis, especially RNA Sequencing, is crucial in research and diagnostics.
• RNA's instability necessitates robust storage and isolation methods to maintain integrity.
• Long-term, cost-effective blood sample storage with minimal volume is desirable for large collections.

Purpose of the Study:

• To assess the feasibility of isolating high-quality RNA from long-term stored blood samples.
• To evaluate a modified RNA isolation protocol for improved RNA integrity.
• To compare RNA quality from long-term stored samples with fresh samples.

Main Methods:

• Blood samples (EDTA and citrate) were stored at -85 °C for 10 years in Boom's lysis buffer.
• RNA was isolated using a modified Zymo Research Quick-RNA kit protocol.
• RNA integrity and quality were assessed, and RNA Sequencing was performed.

Main Results:

• High-quality RNA suitable for transcriptomics was successfully isolated from 10-year-old stored blood samples.
• The modified Zymo Research protocol significantly improved RNA integrity compared to the original Boom method.
• RNA Sequencing yielded high-quality data comparable to studies using recently collected blood.

Conclusions:

• Long-term storage of blood in Boom's lysis buffer at -85 °C preserves RNA for high-quality downstream analysis.
• A modified Zymo Research Quick-RNA kit protocol offers an effective method for isolating RNA from aged samples.
• This approach supports large-scale biobanking and RNA sequencing initiatives.