Rossolini G M, Cresti S, Ingianni A, Cattani P, Riccio M L, Satta G
Dipartimento di Biologia Molecolare, Università di Siena, Italy.
Mol Cell Probes. 1994 Apr;8(2):91-8. doi: 10.1006/mcpr.1994.1013.
The performance of oligonucleotide primers containing deoxyinosine (dl) at all ambiguous positions for polymerase chain reaction, based on ambiguous sequence information derived either from compilations of consensus nucleotide sequences or from amino acid sequences, has been evaluated in two model systems represented respectively by amplification of conserved genomic regions from different types of human papillomavirus and by amplification of a region of the human lysozyme cDNA on the basis of the protein amino acid sequence. In both instances the dl-containing primers obtained the expected amplification products. When using short primers or primers with very high dl contents, however, peculiar reaction conditions had to be adopted to obtain successful amplification and, in the latter case, performance remained suboptimal. Comparison of results with those obtained using corresponding degenerate primers showed that the use of dl-containing primers can be advantageous in terms of both specificity and yield of the amplification product. Sequence analysis of amplification products showed that dG residues are always found at positions corresponding to the dl residues of the primers.
基于来自共有核苷酸序列汇编或氨基酸序列的模糊序列信息，对在所有模糊位置含有脱氧次黄苷（dl）的寡核苷酸引物用于聚合酶链反应的性能，已在两个模型系统中进行了评估，这两个模型系统分别以扩增不同类型人乳头瘤病毒的保守基因组区域以及基于蛋白质氨基酸序列扩增人溶菌酶cDNA的一个区域为代表。在这两种情况下，含dl的引物均获得了预期的扩增产物。然而，当使用短引物或dl含量非常高的引物时，必须采用特殊的反应条件才能获得成功的扩增，并且在后一种情况下，性能仍不理想。将结果与使用相应简并引物获得的结果进行比较表明，使用含dl的引物在扩增产物的特异性和产量方面都可能具有优势。扩增产物的序列分析表明，在与引物的dl残基相对应的位置总是发现dG残基。
