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  Česky / Czech version Klin. Biochem. Metab., 14 (35), 2006, No. 2, p. 96–100.
 
Optimization of procedure for isolation of total RNA from adipose tissue 
Lacinová Z., Dolinková M., Haluzíková D., Krajíčková J., Haluzík M. 

III. interní klinika, 1. LF UK a VFN, Praha
 


Summary:

       Objective: Optimise RNA isolation from fat tissue and verify its parameters. Material and Methods: Adipose tissue (about 100 mg) was collected from patients during surgical operations. Tissue sample was stabilized in RNAlater agent (QIAGEN GmbH, Germany). Total RNA was extracted by following kits: Rneasy Protect Mini, Rneasy Lipid Tissue (QIAGEN GmbH, Germany) and MagNA Pure Compact RNA Isolation (Tissue) for MagNA Pure Compact Instrument (Roche Diagnostics GmbH, Germany). Results: RNA concentrations obtained were as follows: Rneasy Protect Mini Kit 37.5 ± 20.7 µg/ml (N = 63), Rneasy Lipid Tissue Kit 85.9 ± 36.3 µg/ml (N = 29), MagNA Pure Compact RNA Isolation Kit (Tissue) 50.6 ± 13.6 µg/ml (N = 28). R260 nm/280 nm ratios were 1.6–2.0. Comparision of MagNa Pure and QIAGEN RNA isolation by RT-PCR for beta-2- -microglobulin showed similar amplification profiles. Inter– and intra-assay were less than 7%. No DNA contamination of total RNA samples was detected with the exception of samples isolated by Rneasy Protect Mini Kit. Conclusion: Rneasy Lipid Tissue Kit and MagNA Pure Compact RNA Isolation Kit (Tissue) provide RNA samples of high quantity, purity and PCR amplificability. RNA samples are reliable for further processing using methods of molecular biology.

        Key words: RNA isolation, beta-2-microglobulin, RT-PCR.
       

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