畜牧兽医学报2024，Vol.55Issue(8)：3436-3445,10.DOI:10.11843/j.issn.0366-6964.2024.08.017
不同单细胞全基因组扩增体系扩增牛微量血液DNA效果评价
Evaluation of the Effect of Different Single Cell Whole Genome Amplification Systems on the Amplification of Bovine Trace Blood DNA
摘要
Abstract
The purpose of this paper was to evaluate the amplification effect of different single cell whole genome amplification(scWGA)systems on bovine trace blood genomic DNA by next-gen-eration sequencing,and to establish a trace DNA whole genome amplification system.In this study,the whole genome of 1 ng blood genomic DNA of Huaxi cattle was amplified by MDA(multiple displacement amplification)and MALBAC(multiple annealing and looping-based amplification cycles)scWGA systems,and then the sequencing library was constructed based on the amplification products of the two systems and the original undiluted blood DNA.The whole genome sequencing(WGS)was performed using the DNBSEQ-T7RS sequencing platform.The amplification efficiency was evaluated by comparing the fragment's size,concentration and total mass of the amplified products.The amplification effects of the two systems were evaluated by analyzing GC content,average depth,consistent rate of typing and call rate.The results showed that the fragment of the amplified product in MDA system was larger than that in MALBAC system(8 kb vs.0.2-2 kb),the concentration and total mass of the product were significantly higher than those in MALBAC system(P＜0.05).Based on the sequencing data,the genome coverage of MDA system was significantly higher than that of MALBAC system at 1 × and 5 × sequencing depth(P＜0.05).In addition,the consistency of typing rate and call rate of MDA system were significantly higher than MALBAC system,while the allele drop rate and false posi-tive rate of MALBAC system were significantly lower than MALBAC system(P＜0.05).In summary,this study revealed that the characteristics of two amplification systems of MDA and MALBAC were based on the DNA amplification of Huaxi cattle 1 ng blood genome,which provided a theoretical basis for improving the key amplification techniques in Huaxi cattle embryo genome selection and promoting the progress of Huaxi cattle genetics and breeding.关键词
全基因组扩增/MDA/MALBAC/微量DNA扩增体系Key words
whole genome amplification/MDA/MALBAC/trace DNA amplification system分类
农业科技引用本文复制引用
牛一凡,赵学明,李崇阳,杨柏高,张培培,张航,冯肖艺,曹建华,余洲,马友记..不同单细胞全基因组扩增体系扩增牛微量血液DNA效果评价[J].畜牧兽医学报,2024,55(8):3436-3445,10.基金项目
国家重点研发计划政府间重点专项(2022YFE0100200) （2022YFE0100200）
国家自然科学基金国际合作项目(32161143032) （32161143032）
农业农村部和财政部资助:现代农业产业技术体系资助(CARS-36) （CARS-36）
国家家养动物种质资源库、中国农业科学院科技创新工程(ASTIP-IAS06) （ASTIP-IAS06）
