RNA-Integrity and 8-Isoprostane Levels Are Stable in Prostate Tissue Samples Upon Long-Term Storage at −80°C

AbstractWe report the results of a survey of the fishes of Kāne‘ohe Bay, O‘ahu, conducted in 2017 as part of the Smithsonian Institution MarineGEO Hawaii bioassessment. We recorded 109 species in 43 families. The most speciose families were Acanthuridae (11 species), Gobiidae (11 species), Pomacentridae (10) and Chaetodontidae (9 species). Nine of the species that we collected are known or suspected to be introduced to the Hawaiian Islands. Specimens were identified, measured and photographed. All specimen vouchers were fixed in formalin and ultimately transferred to 75% ethanol for long-term storage. For nearly all species, we took multiple tissue samples from specimen vouchers prior to formalin-fixation; we preserved tissues in 95% ethanol and then stored them at −80°C. The 5′-end of the mitochondrial cytochrome oxidase I gene (mtCOI) was sequenced for 94 species to confirm their taxonomic identification. Using these barcode sequences, we also measured genetic distances between collected individuals and their conspecifics from other localities outside Hawaii to verify the hypothesis that Hawaiian populations of species broadly distributed throughout the Indo-Pacific may be genetically distinct. We present select case studies to demonstrate the potential for undiscovered endemism in the Hawaiian fish biota.

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Evaluation of a low-cost procedure for sampling, long-term storage, and extraction of RNA from blood for qPCR analyses

Clinical Chemistry and Laboratory Medicine (CCLM) ◽

10.1515/cclm-2014-1054 ◽

2015 ◽

Vol 53 (8) ◽

Cited By ~ 6

Author(s):

Rasmus B. Mærkedahl ◽

Hanne Frøkiær ◽

Lotte Lauritzen ◽

Stine B. Metzdorff

Keyword(s):

Gene Expression ◽

Clinical Trials ◽

Low Cost ◽

Rna Extraction ◽

Initial Step ◽

Rna Integrity ◽

Long Term Storage ◽

Significant Cost Reduction ◽

Term Storage

AbstractIn large clinical trials, where RNA cannot be extracted immediately after sampling, preserving RNA in whole blood is a crucial initial step in obtaining robust qPCR data. The current golden standard for RNA preservation is costly and designed for time-consuming column-based RNA-extraction. We investigated the use of lysis buffer for long-term storage of blood samples for qPCR analysis.Blood was collected from 13 healthy adults and diluted in MagMAX lysis/binding solution or PAXgene Blood RNA tubes and stored at –20 °C for 0, 1, or 4 months before RNA extraction by the matching method. RNA integrity, yield and purity were evaluated and the methods were compared by subsequent analyses of the gene expression levels ofThe MagMAX system extracted 2.3–2.8 times more RNA per mL blood, with better performance in terms of purity, and with comparable levels of integrity relative to the PAXgene system. Gene expression analysis using qPCR of: The MagMAX system can be used for storage of human blood for up to 4 months and is equivalent to the PAXgene system for RNA extraction. It furthermore, provides a means for significant cost reduction in large clinical trials.

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Long-term storage of tissue samples for cell culture

In Vitro ◽

10.1007/bf02616111 ◽

1978 ◽

Vol 14 (5) ◽

pp. 476-478 ◽

Cited By ~ 7

Author(s):

Harold A. Taylor ◽

Susan E. Riley ◽

Stephen E. Parks ◽

Roger E. Stevenson

Keyword(s):

Cell Culture ◽

Tissue Samples ◽

Long Term Storage ◽

Term Storage

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Quality assessment of RNA in long-term storage: The All Our Families biorepository

PLoS ONE ◽

10.1371/journal.pone.0242404 ◽

2020 ◽

Vol 15 (12) ◽

pp. e0242404

Author(s):

Nikki L. Stephenson ◽

Kylie K. Hornaday ◽

Chelsea T. A. Doktorchik ◽

Andrew W. Lyon ◽

Suzanne C. Tough ◽

...

Keyword(s):

Population Based ◽

Rate Of Change ◽

Population Based Study ◽

Acid Yield ◽

Rna Integrity ◽

Long Term Storage ◽

Term Storage ◽

Over Time

Background The All Our Families (AOF) cohort study is a longitudinal population-based study which collected biological samples from 1948 pregnant women between May 2008 and December 2010. As the quality of samples can decline over time, the objective of the current study was to assess the association between storage time and RNA (ribonucleic acid) yield and purity, and confirm the quality of these samples after 7–10 years in long-term storage. Methods Maternal whole blood samples were previously collected by trained phlebotomists and stored in four separate PAXgene Blood RNA Tubes (PreAnalytiX) between 2008 and 2011. RNA was isolated in 2011 and 2018 using PAXgene Blood RNA Kits (PreAnalytiX) as per the manufacturer’s instruction. RNA purity (260/280), as well as RNA yield, were measured using a Nanodrop. The RNA integrity number (RIN) was also assessed from 5–25 and 111–130 months of storage using RNA 6000 Nano Kit and Agilent 2100 BioAnalyzer. Descriptive statistics, paired t-test, and response feature analysis using linear regression were used to assess the association between various predictor variables and quality of the RNA isolated. Results Overall, RNA purity and yield of the samples did not decline over time. RNA purity of samples isolated in 2011 (2.08, 95% CI: 2.08–2.09) were statistically lower (p<0.000) than samples isolated in 2018 (2.101, 95% CI: 2.097, 2.104), and there was no statistical difference between the 2011 (13.08 μg /tube, 95% CI: 12.27–13.89) and 2018 (12.64 μg /tube, 95% CI: 11.83–13.46) RNA yield (p = 0.2964). For every month of storage, the change in RNA purity is -0.01(260/280), and the change in RNA yield between 2011 and 2018 is -0.90 μ g / tube. The mean RIN was 8.49 (95% CI:8.44–8.54), and it ranged from 7.2 to 9.5. The rate of change in expected RIN per month of storage is 0.003 (95% CI 0.002–0.004), so while statistically significant, these results are not relevant. Conclusions RNA quality does not decrease over time, and the methods used to collect and store samples, within a population-based study are robust to inherent operational factors which may degrade sample quality over time.

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Modelling of Moisture Movement in Wood during Outdoor Storage

Nonlinear Analysis Modelling and Control ◽

10.15388/na.2001.6.1.15210 ◽

2001 ◽

Vol 6 (2) ◽

pp. 3-14 ◽

Cited By ~ 23

Author(s):

R. Baronas ◽

F. Ivanauskas ◽

I. Juodeikienė ◽

A. Kajalavičius

Keyword(s):

Experimental Data ◽

Finite Difference ◽

Climatic Conditions ◽

Two Dimensional ◽

Moisture Movement ◽

Finite Difference Technique ◽

Long Term Storage ◽

Space Formulation ◽

Term Storage

A model of moisture movement in wood is presented in this paper in a two-dimensional-in-space formulation. The finite-difference technique has been used in order to obtain the solution of the problem. The model was applied to predict the moisture content in sawn boards from pine during long term storage under outdoor climatic conditions. The satisfactory agreement between the numerical solution and experimental data was obtained.

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