Mittal Shalu, Bushman Frederic, Orgel Leslie E
The Salk Institute for Biological Studies, La Jolla, California 92037, USA.
J Chem Technol Biotechnol. 2003 Apr;78(4):471-473. doi: 10.1002/jctb.806. Epub 2003 Mar 19.
When [P]-labeled DNA is adsorbed on colloidal gold from a 130mmol dm solution of KHPO, it can subsequently be eluted with cold DNA without undergoing detectable degradation. Similarly, DNA can be incubated in solution in the presence or absence of colloidal gold with high concentrations of -mercaptoethanol or hexane-1-thiol without significant degradation. However, when adsorbed DNA is eluted from gold with solutions of one of the thiols, it is recovered as a mixture of mononucleotides and short oligomers. The extent of degradation increases with increasing concentration of the thiol and with increasing elution temperature. Our results emphasize that in designing protocols involving DNA adsorbed on gold surfaces, it is important to avoid allowing the DNA to come into contact with even moderate concentrations of thiols.
当用130mmol dm的KHPO溶液将[P]标记的DNA吸附到胶体金上时,随后可用冷DNA洗脱,且不会发生可检测到的降解。同样,DNA可在有或没有胶体金存在的溶液中,与高浓度的β-巯基乙醇或己烷-1-硫醇一起孵育,而不会发生明显降解。然而,当用其中一种硫醇溶液从金上洗脱吸附的DNA时,回收的是单核苷酸和短寡聚物的混合物。降解程度随硫醇浓度的增加和洗脱温度的升高而增加。我们的结果强调,在设计涉及吸附在金表面的DNA的实验方案时,重要的是要避免让DNA与即使是中等浓度的硫醇接触。