Pancoska Petr, Moravek Zdenek, Moll Ute M
Department of Pathology, Stony Brook University, New York, NY 11794, USA.
Nucleic Acids Res. 2004 Aug 27;32(15):4630-45. doi: 10.1093/nar/gkh802. Print 2004.
Nucleic acids are molecules of choice for both established and emerging nanoscale technologies. These technologies benefit from large functional densities of 'DNA processing elements' that can be readily manufactured. To achieve the desired functionality, polynucleotide sequences are currently designed by a process that involves tedious and laborious filtering of potential candidates against a series of requirements and parameters. Here, we present a complete novel methodology for the rapid rational design of large sets of DNA sequences. This method allows for the direct implementation of very complex and detailed requirements for the generated sequences, thus avoiding 'brute force' filtering. At the same time, these sequences have narrow distributions of melting temperatures. The molecular part of the design process can be done without computer assistance, using an efficient 'human engineering' approach by drawing a single blueprint graph that represents all generated sequences. Moreover, the method eliminates the necessity for extensive thermodynamic calculations. Melting temperature can be calculated only once (or not at all). In addition, the isostability of the sequences is independent of the selection of a particular set of thermodynamic parameters. Applications are presented for DNA sequence designs for microarrays, universal microarray zip sequences and electron transfer experiments.
核酸是成熟的和新兴的纳米技术所选用的分子。这些技术受益于可容易制造的“DNA处理元件”的高功能密度。为实现所需功能,目前多核苷酸序列是通过一个过程来设计的,该过程涉及针对一系列要求和参数对潜在候选序列进行繁琐费力的筛选。在此,我们提出一种全新的方法,用于快速合理设计大量DNA序列。该方法允许直接对所生成的序列实施非常复杂和详细的要求,从而避免“暴力”筛选。同时,这些序列的解链温度分布狭窄。设计过程的分子部分无需计算机辅助即可完成,通过绘制一个表示所有生成序列的单一蓝图图,采用一种高效的“人工设计”方法。此外,该方法无需进行大量的热力学计算。解链温度只需计算一次(或根本无需计算)。另外,序列的等稳定性与特定热力学参数集的选择无关。文中展示了该方法在微阵列DNA序列设计、通用微阵列拉链序列以及电子转移实验中的应用。