Agarose gel electrophoresis is a routinely employed experimental technique for visualizing nucleic acids, typically combined with ethidium bromide or some other molecular dye staining and a UV transilluminator. However, the utilization of a UV transilluminator is restricted due to high cost and safety considerations. In this study, an inexpensive and simple in-house blue Light Emitting Diode (LED) transilluminator was developed using a high-brightness blue LED, carbon resistor, and smartphone charging cable. Its sensitivity for DNA detection was compared with conventional methods. A detection limit of 4 ng/band was confirmed by observing a 100 bp DNA fragment, which is comparable to ethidium bromide staining and UV transilluminator. Additionally, the in-house blue LED transilluminator can be attached to existing electrophoresis chambers, allowing real-time observation of electrophoresis and offering a superior alternative to conventional methods. In conclusion, the developed in-house blue LED transilluminator demonstrates high sensitivity in detecting DNA fragments and holds potential for advancing low-cost research facilities.