We have reported a one-step method that can enable the growth and parallel patterning of DNA nanofibers on a substrate. This method is based on the processes of solvent vapor-induced buildup and controlled drying front movement and forms parallelly aligned DNA nanofibers exceeding several hundred micrometers in length and 40 nm in diameter on a PDMS sheet. DNA nanofibers initially present on the PDMS sheet were transferred onto another surface using transfer-printing (TP). It was also possible to realize crossed two-dimensional patterns of DNA nanofibers by repeating TP. Polarized fluorescent microscope observations revealed that intercalated dyes are highly ordered in molecular alignment and that DNA strands in the fiber are aligned parallel to the growing direction of fibers.