Evaluating the efficiency of Chelex 100 for DNA extraction in orange (Citrus sinensis)

Abstract

Plant research has recently concentrated on the molecular level, which requires reliable, inexpensive, and fast protocols for extracting DNA. Many researchers have identified Chelex-based DNA extraction methods as having the potential to reduce costs. However, Chelex can also prevent DNA degradation by chelating metal ions. Citrus is one of the top three crops grown globally, with oranges being a well-known species. Therefore, study on the molecular of C. sinensis is essential. This study was conducted in 2022 at the genetic lab of the University of Limpopo- to compare two different extraction methods for DNA from C. sinensis. The DNA from the C. sinensis leaves was extracted using Chelex "method 1" (overnight incubation at 56°C) and "method 2" (Ten-minute incubation at 95°C). A spectrophotometer was employed to measure the extracted DNA's qualitative and quantitative parameters accurately. Furthermore, all samples were subjected to PCR reactions with primers for 28S. The results indicated that "method 2" (1.59-1.84) proved to be the most effective method to obtain high-quality extracted DNA. Additionally, the quantitative and qualitative tests for PCR reaction demonstrated that "method 2" produced better results. In conclusion, although both methods yielded favourable outcomes, "method 2" was ultimately the more successful approach.