Novel buffer for long-term preservation of DNA in biological material at room temperature

The collection and preservation of biological material before DNA analysis is critical for inter alia biomedical research, medical diagnostics, forensics and biodiversity conservation. In this study, we evaluate an in-house formulated buffer called the Forensic DNA Laboratory-buffer (FDL-buffer) for preservation of biological material for long term at room temperature. Human saliva stored in the buffer for 8 years, human blood stored for 3 years and delicate animal tissues from the jellyfish <i>Pelagia noctiluca</i> comb jelly <i>Beroe</i> sp., stored for 4 and 6 years respectively consistently produced high-quality DNA. FDL-buffer exhibited compatibility with standard organic, salting out and spin-column extraction methods, making it versatile and applicable to a wide range of applications, including automation. DNA extractions were performed by Salting out, PCI, ZymoQuick and DNAeazy methods. DNA quantity and quality were assessed using qPCR, Qubit, gel electrophoresis, as well as Sanger sequencing, microsatellite profiling and SNPchip analysis. Preservation of DNA integrity and quality in biological samples is essential in the field of biological science, medicine and forensics. Conventional methods of storage such cold temperature and alcohol fixation can be challenging when sampling in remote destinations or with limited resources. We formulated a novel buffer referred to as the FDL-buffer offering extra-long term (&gt;5 years) preservation of DNA in biological material stored at room temperature. We validated the FDL-buffer on human saliva stored for 8 years, human blood stored for 3 years and delicate animal tissues from the jellyfish <i>Pelagia noctiluca</i> (Scyphozoa Pelagiidae) and comb jelly <i>Beroe</i> sp. (Ctenophora, Beroidae), stored without pre-treatment for 4 and 6 years, respectively. A comparative approach was employed using alternative preservation systems and DNA extraction protocols. The FDL-buffer exceeded the parallel performance of the Oragene OG-500 for preserving saliva, and effectively preserved blood samples, yielding high molecular weight DNA. Successful isolation of high-quality DNA from the marine specimens The purified DNA from the FDL-buffer consistently produced high-quality results in downstream applications, including Sanger sequencing, microsatellite fragment analysis profiling and SNPchip analysis. The FDL-buffer exhibited compatibility with standard organic and salting out extraction methods and commercial spin-column extractions. The FDL-buffer can preserve the gDNA from cells and tissues which allows these biospecimens to be transported and stored successfully without cold storage. The formulation offered a cost effective, versatile and time-saving approach for collection and long-term storage of biological material for DNA analysis.