Primer design

The performance of any marker is very much dependent on the optimal design of the primers. Primers act as specific starter molecules for the amplification of the target of interest. Conventional molecular approaches use only two primers. The OSNA assay uses six different primers which have been specifically designed to avoid the amplification of CK19 pseudogenes or their transcripts, amplification of which would lead to false positive results. Furthermore undesired amplification of genomic DNA is avoided due to precipitation of DNA at low pH during sample preparation and the isothermal reaction temperature of 65°C. Together, these factors contribute towards a more specific and accurate analysis of the LN.