应用DNA—荧光染料法评定瘤胃细菌细胞计数的研究

THE STUDY ON RUMEN BACTERIAL NUMBER COUNT BY DNA FLUORESCENCE DYE

本研究应用瘤胃Succinogenes85细菌 ,用DNA荧光染料法探讨一种新的、简便而快速、无需厌氧操作的方法来评定瘤胃细菌计数。结果表明 ,加进DNA染料后 ,用超声波裂解细胞的时间不能低于30分钟 ,并且裂解后的样品在室温下平衡90分钟 ,对细菌的DNA含量没有影响。应用DNA荧光染料法测定的瘤胃细菌数与用细胞流动器测得的数值相比 ,从统计分析来看 ,没有显著差异(P<0.01)。

In this research,a new,simple,fast,oxygen-free operation method by means of DNA fluorescence dye was investigated in bacterial number count of rumen succinogenes 85 strain.The results showed that for increasing the stability of bacterial DNA content,the time of breaking cell with ultrasonic wave by adding DNA dye cant't be decreased below 30 min,and breaking cell samples should be equilibrated in 90 min under room tempereture.The accuracy and precision of groups added in fluorescence dye in concentration of 4ng/ml were higher than groups in concentration of 2ng/ml.Statistically calculation of bacterial number had been shown there was no significent difference between DNA fluorescence dye and cell flow counter(P<0.01).