用于小鼠遗传质量评价的核酸标准样品的制备与稳定性验证

Preparation and Stability Validation of Nucleic Acid Samples for Genetic Quality Evaluation of Mice

目的制备用于BALB/c、C57BL/6两种小鼠遗传质量评价的核酸标准样品及引物,并对其完整性和稳定性进行验证。方法使用琼脂糖凝胶电泳和紫外分光光度法测定混合核酸标准样品的纯度和浓度。并按照抽样原则进行存放稳定性和运输稳定性验证。存放稳定性以4℃、室温(RT)和37℃这3个温度为观测纵坐标,1、2、3、7和14 d为观测横坐标进行抽样验证;采用保温箱、冰袋和温度计模拟运输条件,分别进行1、3和7 d的运输稳定性验证。结果所制备的BALB/c和C57BL/6两种核酸标准样品A260/A280比值均在1.7~1.9,平均浓度分别为37.53和35.03 ng/μL;稳定性实验表明,两种核酸标准样品及引物在4℃和室温(RT)两种温度下可以稳定存放14 d,且在运输环境中7 d内也能保持稳定。结论BALB/c、C57BL/6两种类型的核酸样品完整性和稳定性良好,可以用于小鼠遗传质量评价。

Objective Two nucleic acid standard samples and primers of BALB/c and C57BL/6 were prepared for genetic quality evaluation of mice,and their integrity and stability were verified.Method Agarose gel electrophoresis and Ultraviolet spectrophotometry were used to determine the purity and concentration of mixed nucleic acid standard samples.And according to the sampling principle,the stability of storage and transportation were verified.The storage stability was verified by sampling at 4℃,room temperature(RT)and 37℃as the ordinate of observation,and 1,2,3,7 and 14 d as the abscissa of observation.An incubator,an ice bag and a thermometer were used to simulate the transportation conditions,and the transportation stability was verified for 1,3 and 7 days respectively.Result The A values of A260/A280 of BALB/c and C57BL/6 standard nucleic acid samples were between 1.7 and 1.9,and the average concentrations were 37.53 and 35.03 ng/μL,respectively.The stability test showed that the two standard nucleic acid samples and primers could be stably stored at 4℃and room temperature(RT)for 14 days,and could also be stably stored in the transportation environment for 7 days.Conclusion BALB/c and C57BL/6 nucleic acid samples have good integrity and stability,which can be used to evaluate the genetic quality of mice.