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

2015 ◽  
Vol 53 (8) ◽  
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.

Susceptibility of Japanese pears to low concentrations of ethylene during storage

2007 ◽  
Vol 47 (12) ◽  
pp. 1480 ◽  
Author(s):  
M. J. Szczerbanik ◽  
K. J. Scott ◽  
J. E. Paton ◽  
D. J. Best
Keyword(s):  
Carbon Dioxide ◽  
Low Cost ◽  
Pyrus Pyrifolia ◽  
Green Colour ◽  
Long Term Storage ◽  
Low Concentrations ◽  
Internal Browning ◽  
Term Storage

The ‘Nijisseiki’ cultivar of Japanese pears (Pyrus pyrifolia) is also known as nashi in Australia. Nashi were exposed to levels of <0.005, 0.01, 0.1 and 1.0 µL/L of ethylene in air during 26 weeks storage at 0°C. Levels of ethylene as low as 0.01 µL/L increased chlorophyll loss and visual green colour. Increasing ethylene levels also increased softening and internal browning, although flesh spot decay was reduced in the presence of ethylene. While it would be worthwhile to remove ethylene during long-term storage of ‘Nijisseiki’ in air, another alternative, adding 2% carbon dioxide to the atmosphere, is suggested as a possible low cost means to overcome the ripening effect of ethylene.

scholarly journals Capture and Purification of Plant Genomic DNA on a Readily-available Cellulose Matrix

2017 ◽  
Author(s):  
Megan M. Thompson ◽  
Estelle M. Hrabak
Keyword(s):  
Arabidopsis Thaliana ◽  
Nucleic Acids ◽  
Plant Species ◽  
Efficient Method ◽  
Genomic Dna ◽  
Low Cost ◽  
Long Term Storage ◽  
Cellulose Matrix ◽  
Term Storage

AbstractWhatman FTA ®Cards are a fast and efficient method for capturing and storing nucleic acids but are cost-prohibitive for some researchers. We developed a method that substitutes readily-available cellulose-based paper and homemade washing buffer for commercial FTA ®Cards and FTA ®Purification Reagent. This method is suitable for long-term storage of DNA from many plant species, including Arabidopsis thaliana, prior to purification and PCR.Method SummaryHere we report a low-cost method for long-term storage of plant genomic DNA on a readily available cellulose matrix.

scholarly journals Long-Term Storage of Small Natural History Specimens Using Gelatin Capsules: A Case Study from the Royal Alberta Museum

2018 ◽  
Vol 32 (1) ◽  
pp. 31-46
Author(s):  
Diana Tirlea ◽  
Carmen Li ◽  
Alwynne B. Beaudoin ◽  
Emily Moffat
Keyword(s):  
Continuous Monitoring ◽  
Low Cost ◽  
Ease Of Use ◽  
Gelatin Capsules ◽  
Long Term Storage ◽  
Pros And Cons ◽  
Storage Methods ◽  
Term Storage

Abstract Museums use gelatin capsules to store small objects and specimens, despite limited documentation of their long-term viability. The Royal Alberta Museum (RAM of Canada) uses gelatin capsules to store seeds, bones, and plant material because of their ease of use, transparency, soft-bodied walls, size availability, and low cost. Recently, RAM staff reported damaged capsules from the palaeontology collections. We evaluated 499 capsules used to store specimens accessioned in 1986 and 1988 and investigated capsule properties using Fourier transform infrared spectroscopy and Oddy testing. Only 4.21% of inspected capsules were dented, cracked, and/or fractured. Based on interviews and testing, we determined that damage to capsules likely resulted during handling (i.e., applied force when opening). We conclude that gelatin capsules offer a good, inexpensive method for long-term storage of small, dried specimens in environmentally controlled conditions. Alternatives to gelatin capsules exist, although their pros and cons require evaluation before use. All storage methods require continuous monitoring for signs of container or specimen deterioration.

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

PLoS ONE ◽  
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.

scholarly journals Long term storage in liquid nitrogen leads to only minor phenotypic and gene expression changes in the mammary carcinoma model cell line BT474

Oncotarget ◽  
2017 ◽  
Vol 8 (21) ◽  
pp. 35076-35087 ◽  
Author(s):  
Judit Fazekas ◽  
Thomas W. Grunt ◽  
Erika Jensen-Jarolim ◽  
Josef Singer
Keyword(s):  
Gene Expression ◽  
Cell Line ◽  
Liquid Nitrogen ◽  
Mammary Carcinoma ◽  
Long Term Storage ◽  
Model Cell ◽  
Term Storage

Low-Cost, Low-Tech Biosolids Treatment via Long-Term Storage: A Comparison of Two Pilot-Scale Studies

2018 ◽  
Vol 2018 (4) ◽  
pp. 1143-1150
Author(s):  
Jennifer G Becker ◽  
Karina Eyre ◽  
Tanner Keyzers ◽  
Christa Meingast ◽  
Eric A Seagren
Keyword(s):  
Low Cost ◽  
Pilot Scale ◽  
Long Term Storage ◽  
Term Storage
Sign in / Sign up

Export Citation Format

Share Document