Kieleczawa J, Dunn J J, Studier F W
Biology Department, Brookhaven National Laboratory, Upton, NY 11973.
Science. 1992 Dec 11;258(5089):1787-91. doi: 10.1126/science.1465615.
When template DNA is saturated with a single-stranded DNA binding protein (SSB), strings of three or four contiguous hexanucleotides (hexamers) can cooperate through base-stacking interactions to prime DNA synthesis specifically from the 3' end of the string. Under the same conditions, priming by individual hexamers is suppressed. Strings of three of four hexamers representing more than 200 of the 4096 possible hexamers primed easily readable sequence ladders at more than 75 different sites in single-stranded or denatured double-stranded templates 6.4 kilobases to 40 kilobase pairs long, with a success rate of 60 to 90 percent. A synthesis of 1 micromole of hexamer supplies enough material for thousands of primings, so multiple libraries of all 4096 hexamers could be distributed at a reasonable cost. Such libraries would allow rapid and economical sequencing. Automating this strategy could increase the speed and efficiency of large-scale DNA sequencing by at least an order of magnitude.
当模板DNA被单链DNA结合蛋白（SSB）饱和时，三或四个连续的六核苷酸（六聚体）串可以通过碱基堆积相互作用协同作用，从而特异性地从该串的3'端引发DNA合成。在相同条件下，单个六聚体的引发作用受到抑制。代表4096种可能六聚体中200多种的三或四个六聚体串，在长度为6.4千碱基至40千碱基对的单链或变性双链模板中的75多个不同位点引发了易于读取的序列阶梯，成功率为60%至90%。1微摩尔六聚体的合成提供了足够用于数千次引发的材料，因此所有4096种六聚体的多个文库可以以合理的成本进行分发。这样的文库将实现快速且经济的测序。将此策略自动化可使大规模DNA测序的速度和效率至少提高一个数量级。
