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Quality Control of RNA Extracted from PAXgene Blood RNA Tubes After Different Storage Periods.

Author(s): Tang R, She Q, Lu Y, Yin R, Zhu P, Zhu L, Zhou M, Zheng C

Publication: Biopreserv Biobank, 2019, Vol. , Page

PubMed ID: 31343263 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   This paper investigated the effects of frozen storage duration and blood clotting on the yield, purity, and integrity of RNA from PAXgene blood. The influence of leukocyte counts on RNA yield was also investigated. Blood was collected from a total of 3724 patients with renal disease between December 2011 and December 2012 in PAXgene tubes and frozen at -80°C. Fifty specimens were randomly selected at each timepoint (1 month, 1 year, 2 years, 3 years, 6 years, and 7 years) and thawed at room temperature. RNA was extracted using the PAXgene Blood RNA Kit and QIAcube and then stored frozen at -80°C. RNA concentration and purity were determined spectrophotometrically. RNA integrity was evaluated by bioanalyzer.

   Summary of Findings:

   The yield of RNA from PAXgene blood was unaffected by the duration of frozen storage (1 month to 7 years) but was very weakly correlated with leukocyte counts (r=0.186, P<0.05). The purity (OD262/280) was significantly higher for RNA from specimens stored for 6 or 7 years than those stored for 1 month to 3 years (P<0.05), but there were no differences in OD260/230 among timepoints. The median RIN value was significantly higher in RNA specimens from blood stored for 1 month to 3 years than in that stored for 6 (P<0.001) or 7 years (P<0.05). RIN was very weakly correlated with RNA yield (r=0.181, P<0.05). The rate of occurrence of RIN ≥ 7.0 was not affected by blood storage duration.

   As expected, RNA from the 19 specimens with blood clots had significantly lower RNA yields and RINs than that from the blood without clots (P<0.0001, both), but OD260/280 was not affected by the presence of blood clots.

   Biospecimens

   • Bodily Fluid - Blood

   Preservative Types

   • PAXgene

   Diagnoses:

   • Other diagnoses

   Platform:

   Analyte	Technology Platform
   RNA	Automated electrophoresis/Bioanalyzer
   RNA	Spectrophotometry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Storage duration	1 month 1 year 2 years 3 years 6 years 7 years
   Biospecimen Aliquots and Components	Blood and blood products	Clotted specimen Non clotted specimen

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