Experimental Analysis of XPCR-based protocols

Abstract

This paper reports some experimental results validating in a broader context a variant of PCR, called XPCR, previously introduced and tested on relatively short synthetic DNA sequences. Basic XPCR technique confirmed to work as expected, to concatenate two genes of different lengths, while a library of all permutations of three different genes (extracted from the bacterial strain Bulkolderia fungorum DBT1) has been realized in one step by multiple XPCR. Limits and potentialities of the protocols have been discussed, and tested in several experimental conditions, by aside showing that overlap concatenation of multiple copies of one only gene is not realizable by these procedures, due to strand displacement phenomena. In this case, in fact, one copy of the gene is obtained as a unique amplification product.