固定化核酸的杂交特性。
Hybridization properties of immobilized nucleic acids.
作者信息
Gingeras T R, Kwoh D Y, Davis G R
出版信息
Nucleic Acids Res. 1987 Jul 10;15(13):5373-90. doi: 10.1093/nar/15.13.5373.
Abstract
The 5'-end attachment of oligonucleotides to dextran supports facilitates the study of the hybridization properties of an immobilized oligonucleotide system. The hybridization properties which were studied include: hybridization capacity and kinetics, hybridization-complex stability, and reagents influencing hybridization efficiency. Results of these experiments reveal that the hybridization efficiencies of support-bound oligonucleotides were 75-80% and 40-50% for single-stranded oligonucleotide targets and long double-stranded targets, respectively. These hybridization efficiencies are dependent upon prehybridizing the support-bound oligonucleotides with dextran sulfate. In addition, comparisons of the relative hybridization efficiencies of the support-bound oligonucleotide and nitrocellulose-based systems have been made which indicate a retention of 13-28% of target sequences on the filters and a detection efficiency of 8-20%.
摘要
将寡核苷酸连接到葡聚糖载体的5′端有助于研究固定化寡核苷酸系统的杂交特性。所研究的杂交特性包括：杂交能力和动力学、杂交复合物稳定性以及影响杂交效率的试剂。这些实验结果表明，对于单链寡核苷酸靶标和长双链靶标，载体结合的寡核苷酸的杂交效率分别为75 - 80%和40 - 50%。这些杂交效率取决于用硫酸葡聚糖对载体结合的寡核苷酸进行预杂交。此外，还对载体结合的寡核苷酸系统和基于硝酸纤维素的系统的相对杂交效率进行了比较，结果表明滤膜上靶序列的保留率为13 - 28%，检测效率为8 - 20%。
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