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- Jain, Pankaj C.; Varadarajan, Raghavan
- Analytical biochemistry 2014 v.449 pp. 90-98
- genes; high-throughput nucleotide sequencing; mutagens; mutants; nucleotide sequences; polymerase chain reaction; thermodynamics
- ... With the development of deep sequencing methodologies, it has become important to construct site saturation mutant (SSM) libraries in which every nucleotide/codon in a gene is individually randomized. We describe methodologies for the rapid, efficient, and economical construction of such libraries using inverse polymerase chain reaction (PCR). We show that if the degenerate codon is in the middle ...
- Su, Yu-Chih; Chen, Hsing-Yin; Ko, Ni Chien; Hwang, Chi-Ching; Wu, Min Hui; Wang, Li-Fang; Wang, Yun-Ming; Chang, Sheng-Nan; Wang, Eng-Chi; Wang, Tzu-Pin
- Analytical biochemistry 2014 v.449 pp. 118-128
- DNA; RNA; fluorescence; fluorescent dyes; fluorescent labeling; melting; polymerase chain reaction
- ... Here we report efficient and selective postsynthesis labeling strategies, based on an advanced phosphoramidation reaction, for nucleic acids of either synthetic or enzyme-catalyzed origin. The reactions provided phosphorimidazolide intermediates of DNA or RNA which, whether reacted in one pot (one-step) or purified (two-step), were directly or indirectly phosphoramidated with label molecules. The ...