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cDNA library construction of two human Demodexspecies

  • DongLing Niu , RuiLing Wang , YaE Zhao EMAIL logo , Rui Yang , Li Hu , YuYang Lei and WeiChao Dan
Published/Copyright: April 18, 2017
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Acta Parasitologica
From the journal Acta Parasitologica Volume 62 Issue 2

Abstract

The research of Demodex, a type of pathogen causing various dermatoses in animals and human beings, is lacking at RNA level. This study aims at extracting RNA and constructing cDNA library for Demodex. First, P. cuniculiand D. farinaewere mixed to establish homogenization method for RNA extraction. Second, D. folliculorumand D. breviswere collected and preserved in Trizol, which were mixed with D. farinaerespectively to extract RNA. Finally, cDNA library was constructed and its quality was assessed. The results indicated that for D. folliculorum& D. farinae, the recombination rate of cDNA library was 90.67% and the library titer was 7.50 × 104 pfu/ml. 17 of the 59 positive clones were predicted to be of D. folliculorum; For D. brevis& D. farinae, the recombination rate was 90.96% and the library titer was 7.85 x104 pfu/ml. 40 of the 59 positive clones were predicted to be of D. brevis. Further detection by specific primers demonstrated that mtDNA cox1, cox3and ATP6 detected from cDNA libraries had 96.52%-99.73% identities with the corresponding sequences in GenBank. In conclusion, the cDNA libraries constructed for Demodexmixed with D. farinaewere successful and could satisfy the requirements for functional genes detection.

Keywords: Demodex; homogenization method; RNA extraction; cDNA library; quality assessment
†

DongLing Niu and RuiLing Wang equally contributed to this paper and should be considered as first co-authors

Acknowledgements

This work was supported by the National Natural Science Foundation of China (grant numbers 81271856 and 81471972).

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Received: 2016-11-16
Revised: 2016-12-20
Accepted: 2017-1-19
Published Online: 2017-4-18
Published in Print: 2017-6-1

© 2017 W. Stefański Institute of Parasitology, PAS

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https://doi.org/10.1515/ap-2017-0043
Keywords for this article
Demodex; homogenization method; RNA extraction; cDNA library; quality assessment

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  4. New record of Steinernema arenarium (Artyukhovsky) (Rhabditida: Steinernematidae) from Ukraine and a note on its distribution
  6. Isolation and molecular characterization of a Naegleria strain from a recreational water fountain in Tenerife, Canary Islands, Spain
  8. PCR denaturing gradient gel electrophoresis as a useful method to identify of intestinal bacteria flora in Haemaphysalis flava ticks
  10. Three new species of Pharygodonidae (Nematoda: Oxyuridea) in Laudakia tuberculata (Squamata: Agamidae) from Dehradun, Uttarakhand, India
  12. Amoebicidal activity of α-bisabolol, the main sesquiterpene in chamomile (Matricaria recutita L.) essential oil against the trophozoite stage of Acanthamoeba castellani Neff
  14. A new species of Oochoristica (Cyclophyllidea: Linstowiidae) from non-native Mediterranean geckos, Hemidactylus turcicus (Sauria: Gekkonidae), from Texas, USA
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  18. Ecological implications of floods on the parasite communities of two freshwater catfishes in a Neotropical floodplain
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