COMPARISON OF METHODS OF DNA EXTRACTION FROM HERMETIA ILLUCENS LARVAE

Abstract

The black soldier fly (Hermetia illucens) is a promising and promising source of animal feed due to its high protein and fat content. For this reason, in 2023, by decision of the Government of the Russian Federation, it was included in the list of agricultural products. Currently, the active use of molecular genetic analysis methods for agricultural purposes continues, including for the study of feed and feed additives. However, today there is too little data in the domestic literature on their use against the black soldier fly. Thus, there is almost no information about DNA extraction methods - the very first stage of any genetic analysis. Thus, the purpose of this study is to compare the effectiveness of existing DNA extraction methods and adapt them to work with Hermetia illucens larvae. In this study, several DNA extraction methods were tested, based on different lysing (SDS, guanidine thiocyanate, CTAB) and chelating (EDTA) agents, as well as lysis durations (1, 2, 3 hours), in comparison with a commercial kit. As a result, it was found that the highest DNA concentration (750 ng/?l) is achieved using the CTAB method, however, when using this protocol, additional purification is necessary. The combined action of SDS and high concentrations of EDTA results in a lower DNA yield (50ng/µl), but does not require additional purification. For the first time, a method based on guanidine thiocyanate was used, which turned out to be quite relevant for this object of study. All of the above methods resulted in comparable or higher DNA yield compared to the commercial GMO-SORB-B kit. Increasing the lysis time to 3 hours when using methods based on guanidine thiocyanate and CTAB leads to increased DNA concentration.