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2018 | 67 | 3 |

Tytuł artykułu

RNA quality control using external standard RNA

Warianty tytułu

Języki publikacji

EN

Abstrakty

EN
In this paper, we propose a new evaluation method using external standard RNA for quality control of the extracted RNA. RNA Integrity Number and UV absorption are generally used as a basis for RNA quality control; however, these methods do not always reflect the quality of mRNA. While standard RNA is supposedly designed on the basis of mRNA, it has the potential to be used to evaluate the quality of the mRNA. In this study, we took into consideration the three essential factors, viz., yield of mRNA, inhibition to DNA polymerase, and degradation of mRNA for determining the RNA quality using standard RNA. It would be possible to know yield of mRNA and inhibition of the enzyme reaction by adding standard RNA before RNA extraction and looking at standard RNA loss. Degradation was evaluated by comparing the differences in the 3’ and 5’ regions of the RNA. In our study, it was demonstrated that in the crude extract of Saccharomyces cerevisiae, degradation was comparatively higher at the 3’ end of RNA than at the 5’ end. Hence, the degree of RNA degradation can be evaluated by comparing the ratio of degradation from the 3’ and 5’ end.

Słowa kluczowe

Wydawca

-

Rocznik

Tom

67

Numer

3

Opis fizyczny

p.347-353,fig.,ref.

Twórcy

autor
  • Faculty of Applied Biological Sciences, Gifu University, Yanagido, Gifu, Japan
autor
  • Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, Tsukuba, Japan
autor
  • Faculty of Applied Biological Sciences, Gifu University, Yanagido, Gifu, Japan

Bibliografia

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  • Cannistraro VJ, Kennell D. 1991. RNase I, a form of RNase I, and mRNA degradation in Escherichia coli. J Bacteriol. 173:4653–4659.
  • Feng H, Zhang X, Zhang C. 2015. mRIN for direct assessment of genome-wide and gene-specific mRNA integrity from large-scale RNA-sequencing data. Nat Commun. 6:7816.
  • Georgiadis AP, Kishore A, Zorrilla M, Jaffe TM, Sanfilippo JS, Volk E, Rajkovic A, Yatsenko AN. 2015. High quality RNA in semen and sperm: isolation, analysis and potential application in clinical testing. J Urol. 193:352–359.
  • Heumüller-Klug S, Sticht C, Kaiser K, Wink E, Hagl C, Wessel L, Schäfer KH. 2015. Degradation of intestinal mRNA: A matter of treatment. World J Gastroenterol. 21:3499–3508.
  • Hui MP, Foley PL, Belasco JG. 2014. Messenger RNA degradation in bacterial cells. Annu Rev Genet. 48:537–559.
  • Imbeaud S, Graudens E, Boulanger V, Barlet X, Zaborski P, Eveno E, Mueller O, Schroeder A, Auffray C. 2005. Towards standardization of RNA quality assessment using user-independent classifiers of microcapillary electrophoresis traces. Nucleic Acids Res. 33:e56.
  • Kashofer K., Viertler C, Pichler M, Zatloukal K. 2013. Quality control of RNA preservation and extraction from paraffin-embedded tissue: implications for RT-PCR and microarray analysis. PLoS One. 8:e70714.
  • Lehnik-Habrink M, Lewis RJ, Mäder U, Stülke J. 2012. RNA degradation in Bacillus subtilis: an interplay of essential endo- and exoribonucleases. Mol Microbiol. 84:1005–1017.
  • Li X, Nair A, Wang S, Wang L. 2015. Quality control of RNA-seq experiments. Methods Mol Biol. 1269:137–146.
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  • Pionzio AM, McCord BR. 2014. The effect of internal control sequence and length on the response to PCR inhibition in real-time PCR quantitation. Forensic Sci Int Genet. 9:55–60.
  • Qin H, Chen X, Tang Y, Hou H, Sheng R, Shen J. 2016. Modified method for the extraction of mRNA from pabby soils. Biotechnol Lett. 38:2163–2167.
  • Sambrook J, Fritsch EF, Maniatis T. 1989. Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press.
  • Szczesny RJ, Borowski LS, Malecki M, Wojcik MA, Stepien PP, Golik P. 2012. RNA degradation in Yeast and Human mitochondria. Biochim Biophys Acta. 1819:1027–1034.
  • Takahashi J, Takatsu A, Iwahashi H. 2013. Evaluation for integrity of extracted RNA by reference material of RNA. J Med Diagn Meth. 2:128.
  • Tong W, Lucas AB, Shippy R, Fan X, Fang H, Hong H, Orr MS, Chu TM, Guo X, et al. 2006. Evaluation of external RNA controls for the assessment of microarray performance. Nat Biotechnol. 24:1132–1139.
  • Wang Z, Gerstein M, Snyder M. 2009. RNA-Seq a revolutionary tool for transcriptomics. Nat Rev Genetics. 10:57–63.

Typ dokumentu

Bibliografia

Identyfikatory

Identyfikator YADDA

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