Comparison of methods for total RNA isolation from aspergillus flavus NSH9

Amylase is a starch digesting enzyme which decomposes starch to yield simple sugar glucose. Due to the commercial value of amylase enzyme, rendering a high demand in the commercial industries. Aspergillus species are the important producer of amylases due to its high enzymes production potential. A very good quality of RNA is critical for gene expression studies. However, many protocols of total RNA extraction are apparently suitable and efficient for certain fungal groups only. In this experiment, the total RNA was extracted from Aspergillus flavus NSH9 by using phenol and LiCl precipitation, hot phenol method with LiCl precipitation and/or NaCl precipitation. These three methods were compared for identifying the suitable and effective protocol to isolate high quality of RNA from Aspergillus flavus NSH9. The result obtained showed the failure extraction of total RNA from Aspergillus flavus NSH9 via phenol and LiCl precipitation. Further experiments proved that it is successfully to isolate high purity and intact RNA from Aspergillus flavus NSH9 by using hot phenol method with LiCl precipitation and/or NaCl precipitation. Both methods produced higher RNA yields with the purity of 1.8 to 2.0 (A260/A280). Therefore, both hot phenol method with LiCl precipitation and/or NaCl precipitation are suitable and effective protocols for extracting RNA from Aspergillus flavus NSH9