SEQUENCE IDENTITY OF THE N-1 PRODUCT OF A SYNTHETIC OLIGONUCLEOTIDE
SEQUENCE IDENTITY OF THE N-1 PRODUCT OF A SYNTHETIC OLIGONUCLEOTIDE
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DOI：
10.1093/nar/23.11.1841
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发表时间：
1995-06-11
影响因子：
14.9
通讯作者：
AGRAWAL, S
中科院分区：
文献类型：
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作者：
TEMSAMANI, J;KUBERT, M;AGRAWAL, S
寡核苷酸合成和纯化后，最终产物通常含有低水平的 n-1 同属物种。我们已经对 25 聚体磷酸二酯寡核苷酸的 n-1 群体进行了测序。从凝胶上切下n-1条带并洗脱。寡核苷酸带有 dA 尾并与带 dT 尾的质粒退火。连接重组质粒并用于转化感受态细菌。我们的结果表明 n-1 群体是异质的。寡聚体3'端截短核苷酸的频率远高于5'端。 5'端最后四个核苷酸未发现截短的核苷酸。我们的结果还表明，寡核苷酸链可以在可控孔径玻璃载体的未反应位点上生长。
After synthesis and purification of an oligonucleotide, the final product usually contains a low level of n-1 congeneric species. We have sequenced the n-1 population of a 25mer phosphodiester oligonucleotide. The n-1 band was cut from the gel and eluted. Oligonucleotides were tailed with dA and annealed to a dT-tailed plasmid. There recombinant plasmid was ligated and used to transform competent bacteria. Our results show that the n-1 population was heterogeneous. The frequency of truncated nucleotides at the 3'-end was much higher than at the 5'-end of the oligomer. No truncated nucleotides were found in the last four nucleotides at the 5'-end. Our results also show that the chain of oligonucleotides can grow on unreacted sites of a controlled-pore glass support.