OPTIMIZATION OF RNA EXTRACTION PROTOCOLS USING DIFFERENT MAIZE TISSUES

Abstract

Five RNA extraction methods were tested (CTAB micro extraction, Concert™ - Invitrogen; Concert™ adapted; TRI Reagent® - Sigma; TRI Reagent® adapted) using two different maize tissues (mesocotyl and root), objectifying to establish an efficient method to extract RNA to be later used in gene expression studies. Concert™ method using the adapted protocol was the most efficient for RNA extraction from both tissues of maize seedlings, originating 2351.35 μg per 100 mg of mesocotyl and 893.75 μg per 100 mg of roots, considering both quantity and quality of the extracted RNA. Thus, samples can be submitted to further steps, like DNAse treatment and cDNA synthesis, for gene expression studies. Results showed that small modifications like change in time and tube positioning during incubation and the use of two extra chloroform washings can improve RNA amount, quality and integrity, from different tissues. These modifications might be used in researches on gene expression.

Keywords

• Nucleic Acids
• Zea mays
• mesocotyl
• root