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Optimization of Breast Biopsy and Mastectomy Sample Collection Procedures for Biobanking, Personalized Medicine, and Research Applications
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Biobanking practice: RNA storage at low concentration affects integrity.

Eloisa Helena Ribeiro Olivieri1, Luana de Andrade Franco, Rafael Gomes Pereira

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Preserving RNA quality is crucial for transcriptomic studies. Storing RNA at -140°C for up to 7 years maintains integrity, but -80°C requires higher concentrations (250 ng/μL) to prevent degradation.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Biobanking

Background:

  • RNA preservation is vital for whole transcriptome analysis in clinical studies.
  • Storage conditions significantly impact RNA quality and downstream applications.
  • Biobanking protocols must ensure RNA integrity for reliable research.

Purpose of the Study:

  • To evaluate RNA quality from head and neck tumors stored at -140°C for up to 7 years.
  • To assess RNA integrity at -80°C under different concentrations and freeze-thaw cycles.
  • To profile the RNA collection quality within the AC Camargo Biobank.

Main Methods:

  • RNA Integrity Number (RIN) was used to measure RNA quality.
  • Tissues stored at -140°C were analyzed after various time intervals.
  • Purified RNA aliquots at 250 ng/μL and 25 ng/μL were stored at -80°C and re-evaluated.
  • RNA from diverse human tumor sites in the biobank was characterized.

Main Results:

  • No significant RNA degradation was observed with storage up to 7 years at -140°C.
  • RNA integrity was maintained at -80°C for 4 years when stored at 250 ng/μL.
  • Significant RNA degradation occurred at 25 ng/μL after 8 months at -80°C.
  • Most biobanked tumor RNAs were high quality (average RIN ~7), with ovary and stomach samples showing degradation.

Conclusions:

  • Optimal RNA preservation requires strict control of both storage temperature and RNA concentration.
  • Long-term storage at -140°C is suitable for maintaining RNA integrity.
  • -80°C storage necessitates higher RNA concentrations to prevent degradation.
  • The AC Camargo Biobank's RNA collection is largely suitable for demanding gene expression analyses due to adherence to SOPs.