Laser-induced fluorescence capillary electrophoresis optimization of random oligonucleotide library of the conditions for polymerase chain reaction amplification.docVIP

 PAGE \* MERGEFORMAT 17 Laser-induced fluorescence capillary electrophoresis optimization of random oligonucleotide library of the conditions for polymerase chain reaction amplification Study: High season Hai-Feng Song Jie Ying Wang Qingqing pay [Abstract] oligonucleotide library by polymerase chain reaction (PCR) amplification is evolution of ligands by exponential enrichment system technology (SELEX) screening aptamer technology to the important steps. In this study, capillary electrophoresis with laser-induced fluorescence detection control (CE LIF), by double-stranded product, PCR-products and remaining primers study, study libraries PCR amplification oligonucleotides influencing factors, and its optimization. The results show that the random oligonucleotide amplified library was amplified with the traditional uniform template significantly different from a dozen of its cycle in the amplification product reached maximum; In addition, the initial number of template, annealing temperature, DNA polymerase concentration conditions are different. So , during the SELEX library must be screened before the detailed PCR conditions were optimized. The library was optimized in the N39 PCR amplification conditions were: initial template of 105 molecules, DNA polymerase concentration of 0.05 U / μL, annealing temperature 70 ℃, 18 cycles. In this study, effective screening for the use of SELEX aptamer technology to reduce false positives and improve screening specificity to provide a reference. [Keywords:] capillary electrophoresis; evolution of ligands by exponential enrichment technology systems; polymerase chain reaction; oligonucleotide library; aptamer 1 Introduction Nucleic acid aptamers (Aptamers) is from a large number of combinatorial library screening of short-chain oligonucleotides, it has a wide range of target molecules, good stability, high affinity characteristics, from the affinity analysis to disease diagnosis and treatment, natural scienc