| 摘要: |
| 采用正交试验设计,从Mg2+、Taq DNA聚合酶、dNTP、微卫星引物和随机引物浓度等方面对朱鹮RMAPD扩增体系进行了优化。结果表明,朱鹮RMAPD扩增的最佳体系为:10×Buffer 2.0 μL,MgCl2 3.13 mmol/L,TaqDNA聚合酶1 U,dNTP 0.10 mmol/L,微卫星引物0.20 μmol/L,随机引物0.50μmol/L,模板DNA 3.75 mg/L,最后补ddH2O至20 μL。将该体系用于朱鹮RMAPD分析,得到了重复性和稳定性良好的朱鹮RMAPD试验结果。 |
| 关键词: 朱鹮 扩增体系 正交试验设计 |
| DOI： |
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| 基金项目:国家自然科学基金项目（No 3007093）；陕西省自然科学基金项目（No 2001SM14）；陕西省林业厅野生动物救护基金项目 |
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| Orthogonal experiment optimization of RMAPD in crested ibis |
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| Abstract: |
| L16(45) orthogonal experiment design was performed with 5 factors of the concentration of Mg2+,Taq DNA polymerase,dNTP and primers for the RMAPD reaction system in crested ibis.The results indicated that the RMAPD reaction system in crested ibis was 10×Buffer 2.0 μL,MgCl2 3.13 mmol/L,Taq DNA polymerase 1 U,dNTP 0.10 mmol/L,microsatellite primer 0.20 μmol/L,random primer 0.50 μmol/L,template DNA 3.75 mg/L,and adding ddH2O to 20 μL.Results of RMAPD in crested ibis with good repeatability and stability were obtained by using the optimized reaction system. |
| Key words: crested ibis(Nipponia Nippon) RMAPD reaction system orthogonal experiment design |
