Improved and convenient method of RNA isolation from polyphenols and polysaccharide rich plant tissues.
It has been difficult to extract a good quality total RNA from the plant parts (such as seeds) which contain high levels of phenolic compounds, carbohydrates and other compounds that bind and/or co-precipitate with RNA. A simple, rapid and efficient method for isolating total RNA from polyphenols and polysaccharide rich plant tissues has been developed. Seeds of leguminosae family were chosen for the study. The good quality and high yield of total RNA was achieved with A260/A280 ratio of 1.9. Seeds of three different crops (Cajanus cajan, Dolichos biflorus and Vigna mungo) at different developmental stages were evaluated for total RNA extraction using standardized protocol. Seeds at 21 days after flowering (DAF) gave the best results among others (7 DAF and dry seeds). Quality of isolated RNA from all the three crops was further checked by cDNA synthesis. The extracted RNA was found suitable for further molecular applications such as reverse transcription and cDNA library construction.
Kansal
Rekha
R
National Research Centre on Plant Biotechnology, Indian Agricultural Research Institute, New Delhi 110 012, India. rekhakansal@hotmail.com
Kuhar
Kalika
K
Verma
Isha
I
Gupta
Ram Niwas
RN
Gupta
Vijay Kumar
VK
Koundal
Kirpa Ram
KR
D005591:Chemical Fractionation / Q000379:methods*; D018076:DNA, Complementary / Q000235:genetics; D007887:Fabaceae / Q000737:chemistry* / Q000235:genetics*; D005419:Flavonoids / Q000032:analysis*; D010636:Phenols / Q000032:analysis*; D059808:Polyphenols; D011134:Polysaccharides / Q000032:analysis*; D018749:RNA, Plant / Q000302:isolation & purification*
D018076:DNA, Complementary; D005419:Flavonoids; D010636:Phenols; D059808:Polyphenols; D011134:Polysaccharides; D018749:RNA, Plant