Comparison of different methods for extraction of viral RNA from avian brain tissue

Podgorski, Iva (2012) Comparison of different methods for extraction of viral RNA from avian brain tissue. Diploma thesis, Faculty of Science > Department of Biology.

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Abstract

Rapid and accurate diagnosis of animal infectious diseases is cruical for timely prevention of the spread and the consequent heavy losses. Molecular biology techniques, especially RNA and DNA isolation, are extensively used in viral infection diagnosis. This study investigates the effect of sample freezing in liquid nitrogen and viral RNA yield by use of four different RNA isolation methods from experimentally infected chickensʼ brain tissue previously stored at -70°C. RNA was isolated by using commercial reagents: 1) Roche High Pure Viral Nucleic Acid Kit, 2) TRIzol®, 3) Qiagen RNeasy Lipid Tissue Mini Kit and 4) combination of TRIzol® and Roche High Pure Viral Nucleic Acid Kit. Yields of isolated RNA were compared by RRT-PCR in LightCycler 1.5. Each method provided enough RNA to detect avian paramyxovirus type 1, but isolation by using Qiagen RNeasy Lipid Tissue Mini Kit and TRIzol® gave the best RNA yield. Freezing of the brain tissue in liquid nitrogen did not influence the yield of RNA.

Item Type: Thesis (Diploma thesis)
Keywords: avian paramyxovirus type 1, Newcastle disease, RRT-PCR
Supervisor: Savić, Vladimir
Co-supervisor: Marijanović, Inga
Date: 2012
Number of Pages: 29
Subjects: NATURAL SCIENCES > Biology
Divisions: Faculty of Science > Department of Biology
Depositing User: Grozdana Sirotic
Date Deposited: 10 Mar 2014 11:18
Last Modified: 19 Sep 2014 12:13
URI: http://digre.pmf.unizg.hr/id/eprint/559

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