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 (A_260/280) 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 A_260/280 values of isolated RNA were 1.984 ± 0.030. DNase treatment further improved the A_260/280 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.

改进的 SDS-Trizol 方法经过优化，可在 4 个月内以一个月的固定间隔从储存的玉米种子中分离总 RNA。在使用 Trizol 之前使用 SDS 提取缓冲液减少了与种子中高碳水化合物含量相关的共沉淀问题。在整个储存间隔期间记录的种子平均 RNA 产量为 978.6 ± 65.46 ng/µl。分离的 RNA 的平均分光光度值 ( A _260/280 ) 从 1.974 ± 0.033 变化到 1.998 ± 0.022。使用 Trizol 方法从绿叶中分离 RNA 的尝试也确保了分离出的 RNA 的质量和数量相当。新鲜叶子的 RNA 产量记录为 1008.2 ± 77.088 ng/µl，略高于几个月种子的平均 RNA 产量。观察到的平均值A _260/280分离的 RNA 的值为 1.984 ± 0.030。DNase 处理进一步提高了种子 (2.003 ± 0.006) 和叶子 (2.012 ± 0.037)的A _260/280比率。RNA提取后的下游分析，如第一链cDNA合成和正常PCR，确保了分离RNA的高质量和数量以及完整性。使用改进的基于 SDS 的 Trizol 方法从储存的种子中提取 RNA，使用 Trizol 方法从新鲜叶子中提取 RNA，为理解涉及发育步骤的关键基因的作用提供了新的可能性，尤其是在储存的种子中。