Method for the design of oligonucleotides for molecular biology techniques

摘要

A method of designing oligonucleotides useful in molecular biology techniques as PCR primers or in other techniques as identification and/or quantification probes is disclosed. The method permits designing specific oligonucleotides for the identification of a determined sequence in a metagenomic sample. The method includes selecting or constructing a database of reference sequences, selecting a subset of sequences belonging to target organisms, selecting candidate oligonucleotides from such sequences, depurating these candidate oligonucleotides according to hybridization specificity and thermodynamic stability criteria, obtaining a list of designed oligonucleotides that fulfill the hybridization specificity and thermodynamic stability criteria, producing materially by chemical synthesis the designed oligonucleotides, and selecting the oligonucleotides which comply with the requirements of the desired process.