The perils of polynucleotides revisited

DNA computing relies on the successful implementation of physical chemistry techniques involving oligonucleotides of prescribed sequence. Our laboratory has been involved in the assembly and manipulation of designed oligonucleotides in order to pursue studies in genetic recombination and nanofabrication. We have constructed a large number of unusual branched DNA motifs used to build a variety of DNA objects, lattices, and nanomechanical devices. Our experience with these systems has uncovered a large number of experimental pitfalls that may confront individuals working with DNA-based computation. A decade ago, we presented our experience in this area in the hope that we could help investigators to anticipate experimental problems that affect DNA computing schemes. Here, we review these points from the vantage point of further experience, indicating both modifications to the original criteria and new points as well.