适配子筛选中随机寡核苷酸文库的构建和PCR条件的优化

Abstract

Abstract: The purpose of this experiment was to construct the oligonucleotide library used in the selection of aptamer and optimize the PCR conditions in the selection process. We constructed the oligonucleotide library and designed primers according to the immobilized sequence of amphi-oligonucleotide using Primer Premier 5.0. The proportion of upstream and downstream primers, quantity of template, anneal temperature and cycle numbers of the symmetric and asymmetric PCR were optimized and their repeatabilities were verified. After optimizing, we set up the best symmetric and asymmetric PCR conditions achieved the desirable dsDNA and ssDNA. The optimized PCR showed high reproducibility. We constructed the oligonucleotide library and optimized PCR condition successfully. This study provided a reference for selection of aptamer and laid a foundation for further study.

Key words: aptamer; oligonucleotide library; PCR; optimization

TIAN Rui-yun, LU Shi-ying, LI Lin, WU Zong-cheng, REN Hong-lin, ZHOU Yu, LI Yan-song, HU Pan, LIU Zeng-shan, WANG Guang-ming. Construction of Oligonucleotide Library in Selection of Aptamer and Optimization of PCR Conditions[J]. China Animal Husbandry & Veterinary Medicine.

References

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Construction of Oligonucleotide Library in Selection of Aptamer and Optimization of PCR Conditions

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