Comparison of rna extraction methods for transcript analysis from the psychrophilic yeast, Glaciozyma antarctica

Izwan Bharudin, N. Z. Zaki, Farah Diba Abu Bakar, N. M. Mahadi, N. Najimudin, R. M. Illias, Abdul Munir Abd. Murad

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

To survive in extremely cold environments, psychrophilic microorganisms produce exopolysaccharides (EPS), which are carbohydrate polymers that constitute a substantial component of the extracellular polymers surrounding cells of these microorganisms. EPS can interfere with RNA extraction and decrease the purity of the RNA extracted from EPS producing microorganisms. In this work, six commercial RNA extraction kits and two published protocols for RNA extraction were evaluated for total RNA extraction from the psychrophilic yeast, Glaciozyma antarctica. All the protocols were optimised to obtain the highest quality of total RNA. The results show that all of the tested commercial kits and the tested conventional methods yielded RNA from G. antarctica, albeit with varying quality. The protocol that utilises TRIzolⓇ reagent was the most effective method for isolating total RNA from G. antarctica of which this protocol resulted in the highest RNA yield and purity compared to other methods. This method of RNA extraction produced RNA of sufficient quality for reverse transcriptase PCR (RT-PCR) to detect the expression of the G. antarctica delta 9-fatty acid desaturase gene as well as for the construction of a G. antarctica cDNA library.

Original languageEnglish
Pages (from-to)71-80
Number of pages10
JournalMalaysian Applied Biology
Volume43
Issue number2
Publication statusPublished - 1 Dec 2014

Fingerprint

Antarctica
Yeasts
RNA
yeasts
exopolysaccharides
methodology
microorganisms
purity
polymers
Polymers
stearoyl-CoA desaturase
Reverse Transcriptase Polymerase Chain Reaction
Gene Library
cDNA libraries
reverse transcriptase polymerase chain reaction
Carbohydrates
carbohydrates

Keywords

  • cDNA library
  • Glaciozyma antarctica
  • Psychrophiles
  • RNA extraction

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)

Cite this

Comparison of rna extraction methods for transcript analysis from the psychrophilic yeast, Glaciozyma antarctica. / Bharudin, Izwan; Zaki, N. Z.; Abu Bakar, Farah Diba; Mahadi, N. M.; Najimudin, N.; Illias, R. M.; Abd. Murad, Abdul Munir.

In: Malaysian Applied Biology, Vol. 43, No. 2, 01.12.2014, p. 71-80.

Research output: Contribution to journalArticle

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