Paper-Based Archiving of Mammalian and Plant Samples for RNA Analysis

Natarajan, Purushothaman; Trinh, Thi Ngoc Diep; Mertz, Lawrence M.; Goldsborough, Mindy D.; Fox, D.K.

doi:10.2144/00296pf01

Cited by 43 publications

(33 citation statements)

References 3 publications

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“…Whatman's FTA paper binds nucleic acids and inactivates ribonucleases but the amounts to be stored and retrieved are low and the storage time is limited. 20 Another procedure uses adsorption of the sample on a solid substrate in the presence of a reducing agent and storage in an organic solvent. 21 However, no validation of this procedure has yet been published.…”

Section: Introductionmentioning

confidence: 99%

An efficient method for long-term room temperature storage of RNA

Fabre

Colotte²,

Luis³

et al. 2013

Eur J Hum Genet

113

110

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RNA is a tool used in many fields, from molecular and cellular biology to medicine and nanotechnology. For most of these uses, the integrity of RNA is required and must be maintained during storage. Even though freezing is currently the storage method of choice, the increasing number of samples to be stored and the costly use of a cold chain have highlighted the need for room temperature preservation methods. Here, we report a new room temperature technology that consists in drying RNA samples in the presence of a stabilizer in stainless steel minicapsules. These air-and water-tight capsules isolate RNA from the atmosphere and maintain an anhydrous and anoxic environment. Through the evaluation of RNA integrity over time at room temperature or 90 1C, we identified atmospheric humidity as a major deleterious factor. The degradation rate dependence in temperature fitted an Arrhenius model, with an activation energy of 28.5 kcal/mol and an extrapolated room temperature degradation rate of 3.2 10 À13 /nt/s (95% confidence interval: 2.3-4.2/nt/s). In these conditions, it is expected that an RNA molecule will be subjected to 0.7-1.3 cut every 1000 nucleotides per century. In addition, we showed that stored RNA is compatible for further analyses, such as reverse transcription-quantitative PCR. No significant change in the C q values was observed over a simulated period of several decades. At last, our data are consistent with a sequence-independent degradation rate of RNA in the solid state.

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Section: Introductionmentioning

confidence: 99%

An efficient method for long-term room temperature storage of RNA

Fabre

Colotte²,

Luis³

et al. 2013

Eur J Hum Genet

113

110

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show abstract

“…An alternative and safe way of transportation of inactivated microorganisms is represented by the Flinders Technology Associates filter paper (FTA † card), which is a chemically treated filter paper designed for the collection and room-temperature storage of biological samples for subsequent analysis (Natarajan et al ., 2000;Rogers & Burgoyne, 2000;Moscoso et al ., 2004;Smith & Burgoyne, 2004). The FTA † cards have been used for multiple molecular studies such as DNA processing from human or wildlife samples (Raina & Dogra, 2002;Smith & Burgoyne, 2004) and recently have become a very interesting approach for the detection of poultry microorganisms, such as Mycoplasmas and infectious bronchitis virus (Moscoso et al ., 2004;Moscoso et al ., 2005 ).…”

Section: Introductionmentioning

confidence: 99%

Use of FTA® filter paper for the molecular detection of Newcastle disease virus

et al. 2006

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“…Furthermore, dried blood spots (DBS) extracted from collection cards have routinely been used for DNA amplification for population screening, e.g., genotyping and for DNA archiving (5,6), but have not been widely used for RT-PCR (7)(8)(9). Such an approach of sample acquisition could similarly be used to measure transcript abundance for nutritionally regulated genes, and, therefore, be of value for nutrient assessment purposes.…”

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confidence: 99%

Zinc supplementation of young men alters metallothionein, zinc transporter, and cytokine gene expression in leukocyte populations

Aydemir

Blanchard

Cousins

2006

Proc. Natl. Acad. Sci. U.S.A.

135

114

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An effective measure to assess zinc status of humans has remained elusive, in contrast to iron, where a number of indicators of metabolism͞function are available. Using monocytes, T lymphocytes, and granulocytes isolated by magnetic sorting and dried blood spots (DBS) derived from 50 l of peripheral blood, we evaluated the response of metallothionein (MT), zinc transporter, and cytokine genes to a modest (15 mg of Zn per day) dietary zinc supplement in human subjects. Transcript abundance was measured by quantitative real-time RT-PCR (QRT-PCR). Zinc supplementation increased MT mRNA abundance by up to 2-fold in RNA from leukocyte subsets, and 4-fold in RNA from DBS. Transcript levels for the zinc transporter genes ZnT1 and Zip3 were increased and decreased, respectively, by zinc supplementation. Expression of the ZnT and Zip genes among leukocyte subsets differ by up to 270-fold. Monocytes and granulocytes from supplemented subjects were activated by LPS, whereas T lymphocytes were activated by mimicking antigen presentation. With zinc consumption, TNF-␣ and IL-1␤ expression was greater in activated monocytes and granulocytes, and IFN-␥ mRNA levels were higher in activated T lymphocytes. These studies show that QRT-PCR is a tool to reliably measure transcript abundance for nutritionally responsive genes in human subjects, and that a small sample of whole dried blood, when appropriately collected, can be used as the source of total RNA for QRT-PCR analysis. The results obtained also show that zinc supplementation of human subjects programs specific leukocytic subsets to show enhanced cytokine expression upon activation by stimulators of immunity.granulocytes ͉ monocytes ͉ nutrition ͉ quantitative RT-PCR ͉ T lymphocytes M ethodological advances to assess gene expression have provided a new spectrum of research tools to identify individual genes and groups of genes that produce normal and altered physiology (1, 2). Quantitative real-time RT-PCR (QRT-PCR) provides a highly sensitive and reproducible method for measuring changes in expression of specific genes through transcript sequence detection. Analytical sensitivity of QRT-PCR rests with the fluorometric basis of this technology, which markedly reduces sample size requirements. The latter makes QRT-PCR an attractive method for clinical, survey, or field studies where the amount of sample is limited.Small amounts of blood, spotted onto filter paper and dried, have been used to examine the blood cell levels of two vitamins (folic acid and retinol) for the purpose of nutritional status assessment of human subjects (3, 4). Furthermore, dried blood spots (DBS) extracted from collection cards have routinely been used for DNA amplification for population screening, e.g., genotyping and for DNA archiving (5, 6), but have not been widely used for . Such an approach of sample acquisition could similarly be used to measure transcript abundance for nutritionally regulated genes, and, therefore, be of value for nutrient assessment purposes. In this regard, we were successful...

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Paper-Based Archiving of Mammalian and Plant Samples for RNA Analysis

Cited by 43 publications

References 3 publications

An efficient method for long-term room temperature storage of RNA

An efficient method for long-term room temperature storage of RNA

Use of FTA® filter paper for the molecular detection of Newcastle disease virus

Zinc supplementation of young men alters metallothionein, zinc transporter, and cytokine gene expression in leukocyte populations

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