Small Amplicons Mutation Library for Vaccine Screening by Error-Prone Polymerase Chain Reaction

• Published in: 东华大学学报（英文版） Vol. 40; no. 2; pp. 171 - 176
• Main Authors: CHENG Manman, ZHANG Yunlong, CHEN Ting, ZHANG Minmin, LU Changrui
• Format: Journal Article
• Language: English
• Published: College of Chemistry,Chemical Engineering and Biotechnology,Donghua University,Shanghai 201620,China 01.04.2023
• Subjects: error-prone polymerase chain reaction; in vitro transcription; DNA library; RNA library
• ISSN: 1672-5220
• DOI: 10.19884/j.1672-5220.202112005

Q523.8; Library construction is a common method used to screen target genes in molecular biology.Most library constructions are not suitable for a small DNA library(＜100 base pair(bp))and low RNA library output.To maximize the library's complexity,error-prone polymerase chain reaction(PCR)was used to increase the base mutation rate.After introducing the DNA fragments into the competent cell,the library complexity could reach 109.Library mutation rate increased exponentially with the dilution and amplification of error-prone PCR.The error-prone PCR conditions were optimized including deoxyribonucleotide triphosphate(dNTP)concentration,Mn2+concentration,Mg2+concentration,PCR cycle number,and primer length.Then,a RNA library with high complexity can be obtained by in vitro transcription to meet most molecular biological screening requirements,and can also be used for mRNA vaccine screening.