Development of a Computer Algorithm for Generation of Primers for Nucleic Acid Sequence Based Amplification (NASBA)

Nucleic acid sequence based amplification (NASBA) is a primer based isothermal method of RNA/DNA amplification. Currently, primer design for NASBA has been restricted to hand creating sequences of oligonucleotides that must follow a set of rules to be compatible for the amplification process. This process of hand-creating primers is prone to error and time intensive. The detection of mutants, post amplification, also offers a benefit in point of care scenarios and the design of hybridization probes for sequences in the region of amplification is also an erroneous and time intensive process. By creating a program to design primers and hybridization probes based on the set of rules provided for a sequence of user input DNA or RNA, one can avoid costly errors in primers design and save time. Utilizing Python (a high-level object-oriented programming language), along with a series of bioinformatic libraries such as Biopython and UNAfold one can definitively choose the best primer sequences for a given sample of DNA.