Extraction of functional total RNA from Cayratia trifolia

Roslina Mat Yazid, Nik Marzuki Sidik

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

RNA extraction is the initial and can be the most critical step in the study of gene expression. Extraction of RNA from Cayratia trifolia plant tissue which contains high levels of polysaccharides, polyphenols and other secondary metabolites is tedious and difficult. We had tested a commercial RNA extraction kit, RNeasy Plant Mini Kit (QIAGEN) and made some modification. The use of both RLT and RLC buffers did not produce any useable RNA. Addition of PEG to the buffers improved RNA yield. However, RLC buffer with PEG 8000 gave better yield compared to RLT buffer with PEG 8000. RNA analysis on agarose gel indicated the presence of 2 clear and distinct 18S and 25S rRNA bands proven that the RNA samples are of high quality. Spectrophotometry analysis on RNA samples showed the OD 260/230 and OD 260/280 ratios were more than 1.8. cDNA display experiments indicated the presence of differentially displayed bands. These results proved that the RNA obtained were functional and of a good quality.

Original languageEnglish
Pages (from-to)3404-3409
Number of pages6
JournalAdvances in Environmental Biology
Volume5
Issue number10
Publication statusPublished - Sep 2011

Fingerprint

Trifolium
RNA
Buffers
buffers
Spectrophotometry
secondary metabolite
Polyphenols
spectrophotometry
polysaccharide
Sepharose
agarose
secondary metabolites
gene expression
Polysaccharides
plant tissues
spectroscopy
polyphenols
polysaccharides
gel
Complementary DNA

Keywords

  • cDNA
  • Differential display
  • Peg 8000
  • rRNA
  • RT-PCR

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Environmental Science(all)

Cite this

Extraction of functional total RNA from Cayratia trifolia. / Yazid, Roslina Mat; Sidik, Nik Marzuki.

In: Advances in Environmental Biology, Vol. 5, No. 10, 09.2011, p. 3404-3409.

Research output: Contribution to journalArticle

Yazid, RM & Sidik, NM 2011, 'Extraction of functional total RNA from Cayratia trifolia', Advances in Environmental Biology, vol. 5, no. 10, pp. 3404-3409.
Yazid, Roslina Mat ; Sidik, Nik Marzuki. / Extraction of functional total RNA from Cayratia trifolia. In: Advances in Environmental Biology. 2011 ; Vol. 5, No. 10. pp. 3404-3409.
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