从含顺式二醇的通用聚合物载体上切割寡脱氧核糖核苷酸的快速条件及其脱保护。
Rapid conditions for the cleavage of oligodeoxyribonucleotides from cis-diol-bearing universal polymer supports and their deprotection.
作者信息
Kumar P, Gupta K C
机构信息
Nucleic Acids Research Laboratory, Centre for Biochemical Technology, Mall Road, Delhi University Campus, Delhi-110 007, India
出版信息
Nucleic Acids Res. 1999 May 15;27(10):e2. doi: 10.1093/nar/27.10.e2.
Two sets of deprotection conditions have been evolved for the deprotection of oligodeoxyribonucleotides and their cleavage from commercially available cis -diol group-bearing universal polymer supports. In the first case, oligodeoxyribonucleotides anchored on the universal support were subjected to one of the standard deprotection conditions followed by treatment with aqueous 0.5 M sodium chloride + 0.2 M sodium hydroxide solution for 30 min at room temperature. In the second case, oligonucleotides bound to the universal support were treated with methanolic sodium hydroxide solution under microwave radiation to obtain fully deprotected oligomers within 4 min. Under both conditions, the cleavage of oligonucleotides from the support and their deprotection occurred quantitatively without any side product formation. The cleaved oligonucleotides were found to be identical in all respects (retention time on HPLC and biological activity in PCR) to the corresponding standard oligo-nucleotides.
已经开发出两组用于寡脱氧核糖核苷酸脱保护及其从市售含顺式二醇基团的通用聚合物载体上裂解的脱保护条件。在第一种情况下，将锚定在通用载体上的寡脱氧核糖核苷酸进行标准脱保护条件之一，然后在室温下用0.5 M氯化钠+0.2 M氢氧化钠水溶液处理30分钟。在第二种情况下，将与通用载体结合的寡核苷酸在微波辐射下用甲醇氢氧化钠溶液处理，以在4分钟内获得完全脱保护的寡聚物。在这两种条件下，寡核苷酸从载体上的裂解及其脱保护均定量发生，没有任何副产物形成。发现裂解的寡核苷酸在所有方面（HPLC上的保留时间和PCR中的生物活性）与相应的标准寡核苷酸相同。
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