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Comparison of three RT-PCR-based methods for relative quantification of mRNA. (CROSBI ID 113340)

Prilog u časopisu | izvorni znanstveni rad | međunarodna recenzija

Breljak, Davorka ; Ambriović-Ristov, Andreja ; Kapitanović, Sanja ; Čačev, Tamara ; Gabrilovac, Jelka Comparison of three RT-PCR-based methods for relative quantification of mRNA. // Food technology and biotechnology, 43 (2005), 4; 379-388-x

Podaci o odgovornosti

Breljak, Davorka ; Ambriović-Ristov, Andreja ; Kapitanović, Sanja ; Čačev, Tamara ; Gabrilovac, Jelka

engleski

Comparison of three RT-PCR-based methods for relative quantification of mRNA.

Comparison of three RT-PCR based methods: semi-quantitative, competitive and real-time RT-PCR for relative quantification of mRNA is presented. Aminopeptidase N expressed on human promyeloid HL-60 cell line, at basal and activated state, served as a model for comparison. HL-60 cells were stimulated with IFN-gamma (6 ng/mL) for 72 h at 37 oC, total cellular RNA was isolated, reverse transcribed to cDNA and semi-quantitative, competitive and real-time RT-PCR were performed to obtain the relative levels of mRNA for aminopeptidase N. The data obtained showed that all three RT-PCR based methods give reliable and comparable results, i.e. approximately two-fold increase of aminopeptidase N mRNA on IFN-gamma stimulated HL-60 cells. Thus, in spite of rapid advances made in the area of real-time RT-PCR, end-point RT-PCR such as competitive and semi-quantitative RT-PCR, although laborious and time consuming, may still remain useful techniques for relative mRNA quantification when small number of samples are to be analyzed.

RT-PCR; mRNA quantification; aminopeptidase N; APN; HL-60

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Podaci o izdanju

43 (4)

2005.

379-388-x

objavljeno

1330-9862

Povezanost rada

Temeljne medicinske znanosti, Biologija

Indeksiranost