Affordable Access

Access to the full text

Two-step method for isolation of high-quality RNA from stored seeds of maize rich in starch

Authors
  • Dutta, Suman1
  • Muthusamy, Vignesh1
  • Chhabra, Rashmi1
  • Zunjare, Rajkumar U.1
  • Hossain, Firoz1
  • 1 ICAR-Indian Agricultural Research Institute, New Delhi, 110012, India , New Delhi (India)
Type
Published Article
Journal
3 Biotech
Publisher
Springer International Publishing
Publication Date
Sep 16, 2020
Volume
10
Issue
10
Identifiers
DOI: 10.1007/s13205-020-02424-w
Source
Springer Nature
Keywords
License
Yellow

Abstract

A modified SDS–Trizol method was optimized for isolation of total RNA from the stored maize seeds at regular interval of one month for 4 months. Use of SDS extraction buffer before the use of Trizol reduced the co-precipitation problem associated with high carbohydrate content in the seed. Recorded mean RNA yield from seeds across the storage intervals was 978.6 ± 65.46 ng/µl. Average spectrophotometric values (A260/280) of isolated RNA varied from 1.974 ± 0.033 to 1.998 ± 0.022. Attempts to isolate RNA from green leaves using Trizol method also ensured comparable quality and quantity of the isolated RNA. RNA yield from fresh leaves was recorded 1008.2 ± 77.088 ng/µl which is slightly higher than the mean RNA yield from seeds across months. Observed mean A260/280 values of isolated RNA were 1.984 ± 0.030. DNase treatment further improved the A260/280 ratio in both seeds (2.003 ± 0.006) and leaves (2.012 ± 0.037). High quality and quantity along with integrity of the isolated RNA was ensured through downstream analysis after RNA extraction such as first-strand cDNA synthesis and normal PCR. Extraction of RNA from the stored seeds using modified SDS-based Trizol method and from fresh leaves using Trizol method opened new possibility of understanding role of key genes involving developmental steps especially in the stored seeds.

Report this publication

Statistics

Seen <100 times