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Biochemistry (Moscow)

, Volume 79, Issue 7, pp 706–716 | Cite as

MS2 phage ribonucleoproteins as exogenous internal control for RT-qPCR data normalization in gene expression study of developing rat brain

  • L. A. FedoseevaEmail author
  • O. B. Shevelev
  • N. G. Kolosova
  • G. M. Dymshits
Article

Abstract

The most popular strategy for normalization of RT-qPCR data involves presenting them in comparison with expression of “housekeeping” genes. However, the required stable expression of the control genes is not always achievable. As an alternative, we used ribonucleoprotein phage particles as an exogenous internal control and demonstrated that this type of normalization provides a simple and reliable method for quantification in RT-qPCR experiments. Using phage-based normalization, we analyzed mRNA levels of three popular housekeeping genes coding β-actin, glyceraldehyde-3-phosphate dehydrogenase, and ribosomal protein L30 and showed high variability in their expression patterns during rat brain development, indicating that they should not be used as controls in gene expression studies of the developing brain either individually or in combination. Using phage-based controls, we showed interstrain differences and age-related changes in the expression of genes involved in proteoglycan biosynthesis and degradation in developing brain of senescenceaccelerated OXYS rats and control Wistar rats.

Key words

PCR data normalization reference genes housekeeping genes exogenous internal control ribonucleoprotein particles senescence-accelerated OXYS rats 

Abbreviations

Actb

β-actin gene

EIAC

exogenous internal amplification control

Ext1 and Ext2

exostosins (EXT) 1 and 2 genes, respectively

Gapdh

glyceraldehyde-3′-phosphate dehydrogenase gene

Hpse

heparanase gene

HSPG

heparan sulfate proteoglycans

Rpl30

ribosomal protein L30 gene

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Copyright information

© Pleiades Publishing, Ltd. 2014

Authors and Affiliations

  • L. A. Fedoseeva
    • 1
    Email author
  • O. B. Shevelev
    • 1
  • N. G. Kolosova
    • 1
  • G. M. Dymshits
    • 2
  1. 1.Institute of Cytology and GeneticsSiberian Branch of the Russian Academy of SciencesNovosibirskRussia
  2. 2.Novosibirsk State UniversityNovosibirskRussia

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