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| Purity Analysis of Oligonucleotides Based on Capillary Gel Electrophoresis |
| WANG Le-le1,ZHANG Xiao-xia2,LIU Gang1 |
1.Key Laboratory of Bioanalysis and Metrology for State Marker Regulation, Shanghai Institute of Measurement and Testing Technology, Shanghai 201203, China
2. SCIEX, Beijing 100015, China |
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Abstract During the chemical synthesis of oligonucleotides, the key technical challenge at present is that the full-length sequence is generated with single (n-1) and double (n-2) deletions as the major impurities. Due to the only subtle difference between the molecular mass and charge properties of impurities and the full-length sequence, it is difficult to accurately quantify the purity of oligonucleotides. With the advantages of small sample amount, fast speed, high sensitivity, separation effective, and high degree automatization, capillary gel electrophoresis is widely used in the separation and analysis of nucleic acids. Capillary gel electrophoresis was used to analyze the purity of oligonucleotides, which can effectively separate the full-length sequence of oligonucleotides and n-1 impurities. The linearity is good in the concentration range of 0.03 mg/L to 1.00mg/L (R2=0.9988), and the detection limit is about 0.01mg/L (S/N>3).The establishment of the aboved method will provide strong technical support for the purity analysis and quality control of oligonucleotides.
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Received: 05 May 2022
Published: 08 March 2023
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2023, Vol. 44 
