Laboratoire de Génétique Moléculaire des Eucaryotes du CNRS, Unité 184 de Biologie Moléculaire et de Génie Génétique de 1'INSERM, Faculté de Médecine, 11 Rue Humann, 67085 Strasbourg Cedex, France.
EMBO J. 1984 Apr;3(4):801-5. doi: 10.1002/j.1460-2075.1984.tb01888.x.
An inexpensive, extremely rapid manual method for simultaneous synthesis of large numbers of oligodeoxyribonucleotides on 50 or 150 nanomole scale is described. The oligonucleotides are assembled in parallel by the phosphotriester method on small cellulose paper disks in a simple gas pressure-controlled continuous-flow system. For each addition of a nucleotide the disks are sorted into four sets which are placed in four columns for addition of A, C, G and T, respectively. During one 2-week period, three rounds of synthesis by this method yielded 254 oligomers (8- to 22-mers), many of which were also purified during this time. Using 50 nanomole scale reactions the yields for 17-mers, for example, were in the range of 0.5 O.D.(260) units ( 5 nmol, i.e., 10% yield), an amount sufficient for most purposes. The equipment required is relatively inexpensive and for the most part usually already available in molecular biology laboratories. All chemicals are commercially available and the current reagent cost per oligonucleotide (25 mug, average length 17-mer) is 3 US dollars.
描述了一种在 50 或 150 毫摩尔规模下同时合成大量寡脱氧核苷酸的廉价、极快速的手动方法。寡核苷酸通过磷酸三酯法在小纤维素纸盘上平行组装，在简单的气压控制连续流动系统中进行。对于每个核苷酸的添加，将纸盘分成四组，分别放置在四个列中，用于添加 A、C、G 和 T。在两周的时间内，通过这种方法进行了三轮合成，得到了 254 个寡聚物（8 至 22 个核苷酸），其中许多在这段时间内也得到了纯化。例如，使用 50 毫摩尔规模的反应，17 个核苷酸的产率在 0.5 O.D.（260）单位（5 nmol，即 10%的产率）范围内，足以满足大多数目的。所需的设备相对便宜，而且在大多数情况下，分子生物学实验室通常已经具备了这些设备。所有化学物质均可商购，每个寡核苷酸的当前试剂成本（25 微克，平均长度为 17 个核苷酸）为 3 美元。
