Preloaded PCR tubes for ready-to-use and ambient storage

Abstract

This invention relates to the field of molecular biology and nucleic acid diagnostics: specifically, compositions and methods to facilitate convenient, error- proof, reliable, portable and cost-effective PCR reactions. This universal PCR kit is suitable for use in clinical diagnostics, pathogen detection, genetic screening, forensic testing, and research laboratories, particularly in resource-limited settings or wherever cold-chain maintenance is impractical or difficult to maintain. The present invention provides ready-to-use, storeable at room temperature PCR tubes including master mixes containing all essential components for polymerase chain reaction (PCR) amplification. Each tube is preloaded with an indigenously prepared Taq polymerase enzyme, PCR buffer, dNTPs, MgCl2, and target-specific primers (only in case of customized PCR kits) in a dried formulation for extended shelf-life. This invention aims to reduce cold-chain requirements, minimize handling and preparation errors, streamline diagnostic and research workflows. Conventional PCR workflows require multiple liquid reagents stored under refrigeration or freezing conditions. Preparing each reaction requires multiple and precise aliquoting which is time-consuming specially when dealing with large sample numbers and prone to errors and contamination due to multiple pipetting and repeated actions. While commercial lyophilized PCR kits exist, they typically rely on expensive imported enzymes, require lengthy procurement procedures, and several handling / rehydration steps involving extensive cold chain and difficult to maintain lab resources. The present invention has a locally produced Taq polymerase and a novel drying process that confers extended stability at ambient temperatures, eliminating the need for ultra-low temperature shipping and storage. The ready to use nature in single tube format, room temperature storage / shipment and availability at comparatively low cost makes this technology a viable product for commercialization with ideal solution for biology work having bright commercial potential.