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Comparison of two RNA isolation methods' for determination of SOD1 and SOD2 gene expression in human blood and mononuclear cells

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2013
Comparison_of_two_pub_2013.pdf (97.90Kb)
Authors
Vujović, Ana
Spasojević-Kalimanovska, Vesna
Bogavac-Stanojević, Nataša
Spasić, Slavica
Kotur-Stevuljević, Jelena
Jelić-Ivanović, Zorana
Article (Published version)
Metadata
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Abstract
In the current study, two RNA isolation techniques were compared and their abilities to produce high-quality RNA were evaluated. mRNA expression profiles of SOD1 (Cu/Zn superoxide dismutase) and SOD2 (Mn superoxide dismutase) genes were measured by real-time PCR. From a pool of fresh human citrate-whole blood and ten healthy individuals, RNA was isolated with the TRIzol (TM) extraction method (TRI) and with the ABI PRISM (TM) 6100 Nucleic AcidPrepStation (ABI). The concentration and purity of RNA extracts were determined spectrophotometrically. RNA integrity was evaluated by electrophoresis on a 1% agarose gel. PCR was performed on a 7500 Real-Time PCR System. The student's t-test was applied to compare normally distributed variables. Both protocols gave similar RNA quantities when adjusted to the initial blood volume. Relative quantification values obtained from the TRI method for SOD1 were significantly higher (p lt 0.01) and for SOD2 were significantly lower (p lt 0.05) as compared ...to those obtained from the ABI method, respectively. Coefficients of variation (CV) for gene expression parameters in SOD1 and SOD2 analyses were lower when the TRI method was used. The TRI method was generally more consistent in yielding pure RNA in comparison to the ABI and better reproducibility in gene expression analyses was apparent.

Keywords:
AbiPrism (TM) / blood / PBMC / real-time PCR / SOD / TRIzol (TM)
Source:
Indian Journal of Biotechnology, 2013, 12, 4, 468-474
Publisher:
  • Natl Inst Science Communication-Niscair, New Delhi
Funding / projects:
  • Interactive role of dyslipidemia, oxidative stress and inflammation in atherosclerosis and other diseases: genetic and biochemical markers (RS-175035)
  • The study was supported by COST BM904.

ISSN: 0972-5849

WoS: 000332435700004

Scopus: 2-s2.0-84893396686
[ Google Scholar ]
10
8
Handle
https://hdl.handle.net/21.15107/rcub_farfar_1945
URI
https://farfar.pharmacy.bg.ac.rs/handle/123456789/1945
Collections
  • Radovi istraživača / Researchers’ publications
Institution/Community
Pharmacy
TY  - JOUR
AU  - Vujović, Ana
AU  - Spasojević-Kalimanovska, Vesna
AU  - Bogavac-Stanojević, Nataša
AU  - Spasić, Slavica
AU  - Kotur-Stevuljević, Jelena
AU  - Jelić-Ivanović, Zorana
PY  - 2013
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/1945
AB  - In the current study, two RNA isolation techniques were compared and their abilities to produce high-quality RNA were evaluated. mRNA expression profiles of SOD1 (Cu/Zn superoxide dismutase) and SOD2 (Mn superoxide dismutase) genes were measured by real-time PCR. From a pool of fresh human citrate-whole blood and ten healthy individuals, RNA was isolated with the TRIzol (TM) extraction method (TRI) and with the ABI PRISM (TM) 6100 Nucleic AcidPrepStation (ABI). The concentration and purity of RNA extracts were determined spectrophotometrically. RNA integrity was evaluated by electrophoresis on a 1% agarose gel. PCR was performed on a 7500 Real-Time PCR System. The student's t-test was applied to compare normally distributed variables. Both protocols gave similar RNA quantities when adjusted to the initial blood volume. Relative quantification values obtained from the TRI method for SOD1 were significantly higher (p lt 0.01) and for SOD2 were significantly lower (p lt 0.05) as compared to those obtained from the ABI method, respectively. Coefficients of variation (CV) for gene expression parameters in SOD1 and SOD2 analyses were lower when the TRI method was used. The TRI method was generally more consistent in yielding pure RNA in comparison to the ABI and better reproducibility in gene expression analyses was apparent.
PB  - Natl Inst Science Communication-Niscair, New Delhi
T2  - Indian Journal of Biotechnology
T1  - Comparison of two RNA isolation methods' for determination of SOD1 and SOD2 gene expression in human blood and mononuclear cells
VL  - 12
IS  - 4
SP  - 468
EP  - 474
UR  - https://hdl.handle.net/21.15107/rcub_farfar_1945
ER  - 
@article{
author = "Vujović, Ana and Spasojević-Kalimanovska, Vesna and Bogavac-Stanojević, Nataša and Spasić, Slavica and Kotur-Stevuljević, Jelena and Jelić-Ivanović, Zorana",
year = "2013",
abstract = "In the current study, two RNA isolation techniques were compared and their abilities to produce high-quality RNA were evaluated. mRNA expression profiles of SOD1 (Cu/Zn superoxide dismutase) and SOD2 (Mn superoxide dismutase) genes were measured by real-time PCR. From a pool of fresh human citrate-whole blood and ten healthy individuals, RNA was isolated with the TRIzol (TM) extraction method (TRI) and with the ABI PRISM (TM) 6100 Nucleic AcidPrepStation (ABI). The concentration and purity of RNA extracts were determined spectrophotometrically. RNA integrity was evaluated by electrophoresis on a 1% agarose gel. PCR was performed on a 7500 Real-Time PCR System. The student's t-test was applied to compare normally distributed variables. Both protocols gave similar RNA quantities when adjusted to the initial blood volume. Relative quantification values obtained from the TRI method for SOD1 were significantly higher (p lt 0.01) and for SOD2 were significantly lower (p lt 0.05) as compared to those obtained from the ABI method, respectively. Coefficients of variation (CV) for gene expression parameters in SOD1 and SOD2 analyses were lower when the TRI method was used. The TRI method was generally more consistent in yielding pure RNA in comparison to the ABI and better reproducibility in gene expression analyses was apparent.",
publisher = "Natl Inst Science Communication-Niscair, New Delhi",
journal = "Indian Journal of Biotechnology",
title = "Comparison of two RNA isolation methods' for determination of SOD1 and SOD2 gene expression in human blood and mononuclear cells",
volume = "12",
number = "4",
pages = "468-474",
url = "https://hdl.handle.net/21.15107/rcub_farfar_1945"
}
Vujović, A., Spasojević-Kalimanovska, V., Bogavac-Stanojević, N., Spasić, S., Kotur-Stevuljević, J.,& Jelić-Ivanović, Z.. (2013). Comparison of two RNA isolation methods' for determination of SOD1 and SOD2 gene expression in human blood and mononuclear cells. in Indian Journal of Biotechnology
Natl Inst Science Communication-Niscair, New Delhi., 12(4), 468-474.
https://hdl.handle.net/21.15107/rcub_farfar_1945
Vujović A, Spasojević-Kalimanovska V, Bogavac-Stanojević N, Spasić S, Kotur-Stevuljević J, Jelić-Ivanović Z. Comparison of two RNA isolation methods' for determination of SOD1 and SOD2 gene expression in human blood and mononuclear cells. in Indian Journal of Biotechnology. 2013;12(4):468-474.
https://hdl.handle.net/21.15107/rcub_farfar_1945 .
Vujović, Ana, Spasojević-Kalimanovska, Vesna, Bogavac-Stanojević, Nataša, Spasić, Slavica, Kotur-Stevuljević, Jelena, Jelić-Ivanović, Zorana, "Comparison of two RNA isolation methods' for determination of SOD1 and SOD2 gene expression in human blood and mononuclear cells" in Indian Journal of Biotechnology, 12, no. 4 (2013):468-474,
https://hdl.handle.net/21.15107/rcub_farfar_1945 .

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