| 样本数量和终质量浓度对DNA混合池PCR扩增质量和测序效果的影响 |
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| 引用本文：徐建峰，容维中，石福岳，王燕燕，董 和，高 博，张艳丽，李振明.样本数量和终质量浓度对DNA混合池PCR扩增质量和测序效果的影响[J].西北农业学报,2019,28(5):681~685 |
| DOI:10.7606/j.issn.1004-1389.2019.05.001 |
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| 基金项目:甘肃省重点研发计划-农业类（17YF1NL052）。 |
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| 中文摘要:为筛选适宜的DNA混合池法，采用两因素交叉分组设计，以混合样本数量（20、60和150个）和终质量浓度（50、100 和150 ng/μL）为组合因子，分析不同混合池对PCR扩增质量及SNPs检出率的影响。结果表明：在同一混合样本数量下，以终质量浓度100 ng/μL建立的DNA混合池扩增的PCR产物纯度和 OD260/OD280均优于50 ng/μL，但对测序结果无显著影响；以终质量浓度150 ng/μL建立的DNA混合池扩增的PCR产物杂带较多，不符合测序要求。在同一混合样本终质量浓度下，混合样本数越多，SNPs检出率越高。综上，从PCR扩增质量、SNPs检出率和试验成本来分析，以混合样本150个、终质量浓度100 ng/μL组合建立的DNA混合池效果最好。 |
| 中文关键词:DNA混合池 样本数量 终质量浓度 早胜牛 PCR扩增 SNPs |
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| Effects of Sample Numbers and Final Mass Concentration on PCR Amplification Quality and Sequencing Result with DNA Mixing Pool |
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| Abstract:To screen suitable method for DNA mixing pool, the experimental design of two-factor cross-sectional with mixed sample numbers（20, 60 and 150）and final mass concentration（50,100 and 150 ng/μL）were used to analyze the effect of different DNA mixing pools on PCR amplification quality and SNPs detection efficiency.The results indicated that under the same mixed sample numbers, the PCR products purity and OD260/OD280 amplified by DNA mixing pool of 100 ng/μL final mass concentration were better than 50 ng/μL final mass concentration, and their sequencing results had no significant effect. PCR amplification product with DNA mixing pool of 150 ng/μL final mass concentration didn’t meet sequencing requirements because it had more heterotopic bands. The more mixed sample numbers, the higher SNPs detection efficiency under the same final mass concentration. In conclusion, the DNA mixing pool for the combination of 150 mixed samples and 100 ng/μL final mass concentration was the best. |
| keywords:DNA mixing pool Sample numbers Final mass concentration Zaosheng cattle PCR amplification SNPs |
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