In situ Synthesis of Oligonucleotide and Detection of Gold-label-silver-stain on PTFE Slices  

作　　者：谭美军 汤建新 陈洪 

机构地区：[1]Key Laboratory of Green Packaging and Biological Nanotechnology of Hunan Province, Hunan Polytechnic University, Zhuzhou, Hunan 412008, China [2]Key Laboratory of Non-timber Breeding and Cultivation of Ministry of Forestry of China, Central South Forestry University, Zhuzhou, Hunan 412006, China

出　　处：《Chinese Journal of Chemistry》2005年第9期1247-1252,共6页中国化学（英文版）

基　　金：Project supported by Hunan Province Natural Science Foundation （No. 04jj40023）, the Trans-century Training Program Foundation for the Talents by the Ministry of Education of China, and the Post-doctoral Science Foundation of China.

摘　　要：Poly（tetrafluoroethylene） （PTFE） was treated with plasma in a mixture of nitrogen and hydrogen （1 ： 2 in volume）. X-ray photoelectron spectroscopy （XPS） demonstrated the success of amino group grafting. The as-treated PTFE slices were successively applied to the in situ synthesis of oligonucleotides. With the detection of gold-label-silver-stain, the hybridization signals were recorded with a gel document and analysis system. A target DNA concentration as low as 10 pmol/L could be detected. The complementary and mismatched sequences were distinguished clearly, and the ratio of background-subtracted gray scale values for perfect match ： 1 base mismatch ： 2 base mismatch ： 3 base mismatch was 72 ： 44.4 ： 22.5 ： 11.4. The sensitivity of in situ synthesis system was 1 order of magnitude higher than that of spotting system, and the signal of the former was about 1.5 times stronger than that of the latter under the same target DNA concentration. These plasma modified PTFE slices might open novel prospects for the in situ synthesis of DNA micro-arrays.Poly（tetrafluoroethylene） （PTFE） was treated with plasma in a mixture of nitrogen and hydrogen （1 ： 2 in volume）. X-ray photoelectron spectroscopy （XPS） demonstrated the success of amino group grafting. The as-treated PTFE slices were successively applied to the in situ synthesis of oligonucleotides. With the detection of gold-label-silver-stain, the hybridization signals were recorded with a gel document and analysis system. A target DNA concentration as low as 10 pmol/L could be detected. The complementary and mismatched sequences were distinguished clearly, and the ratio of background-subtracted gray scale values for perfect match ： 1 base mismatch ： 2 base mismatch ： 3 base mismatch was 72 ： 44.4 ： 22.5 ： 11.4. The sensitivity of in situ synthesis system was 1 order of magnitude higher than that of spotting system, and the signal of the former was about 1.5 times stronger than that of the latter under the same target DNA concentration. These plasma modified PTFE slices might open novel prospects for the in situ synthesis of DNA micro-arrays.

关 键 词：OLIGONUCLEOTIDE gold-label-silver-stain in situ synthesis plasma poly（tetrafluoroethylene） 

分 类 号：O629.74[理学—有机化学]