Polymerase chain reaction (PCR) is a valuable tool in the field of molecular biology and molecular diagnostics. PCR using human DNA obtained whole blood can be applied to identify genetic diseases and polymorphisms, or detect microbial infections. However, DNA isolation from various biospecimens such as whole blood, is a laborious and sample-consuming step, and hampers the use of PCR as automatic procedure for large-scale diagnosis. In direct amplification, numerous molecules in biospecimens inhibit the amplification reaction.
We have developed a novel buffer system ‘AnyDirect’ that conserves the enzymatic activity of DNA polymerases for effective use in direct PCR. The amplification of nucleic acids from biospecimens was achieved by using this solution to overcome strong inhibitors of PCR. And the solution could be performed the gene amplification from blood with various thermostable DNA polymerases without DNA isolation. Importantly, low copy number DNA in blood was effectively amplified. Using direct amplification from biospecimens without DNA isolation, we may obtain the many benefits such as time savings, convenience, avoidance of infection in sample handlers, prevention of loss of trace samples in the DNA purification step, and potential for large-scale diagnosis.
In hard tissue and plant, a simple lysis system that could produce an appropriate template for direct PCR with AnyDirect PCR buffer are made possible the direct amplification of DNA fragments from these tissues. Thus, this procedure provides a simple, convenient, non-hazardous, inexpensive, and rapid process for the amplification of DNA from hard tissues.
AnyDirect buffer allows direct PCR from biospecimens and may facilitate detection of genetic diseases or infections by eliminating the time and effort for DNA extraction. The direct PCR with AnyDirect may generate high-throughput results for large-scale diagnosis, investigation of various medical conditions, or application for use with lab-on-a-chip technology.