高效液相色谱与荧光法测定细菌染色体碱基组成的比较

目的 比较连续荧光检测法（荧光法）与高效液相色谱（ＨＰＬＣ）法测定细菌染色体鸟嘌呤胞嘧啶百分比（ＧＣ％）的稳定性、准确性和适应性。方法 应用双股ＤＮＡ结合染料和能够快速速升降温度的连续荧光检测仪、测定９株细菌染色体的变性温度℃值，并根据相应的公式算出ＧＣ％：Ｃ ＨＰＬＣ法直接检测细菌染色体的ＧＣ％。结果 ＤＮＡ纯度对荧光法测得的ＧＣ％结果无显著影响，但影响ＨＰＬＣ的测定结果，荧光法３次检测获得的９

Light cycler and high pressure liquid chromatography in determing the guanine plus cytosine content of bacterial chromosome DNA

Objective To compare stability, accuracy and suitability of Light Gycler and high prescure liquid chromatography in detecting mole percent guanine plus cytosine content (GC%).Methods Light Cycler method, was used following the melting curves and the values of Tm were obtained from the computer screen of Light Cycler, to calculate the GC% according to the formula: GC%x=GC%r - 8.567 (Tmx Tmr) 0.066 (Tmx 2 Tmr 2). The GC% of bacterial chromosome DNA was obtained directly by HPLC within 30 min after the samples were loaded. Results By Light Cycler we detected the values of the Tm from 24 samples obtained within 10 min. The purity of DNA was not visiblely affect the result. The GC% from HPLC method was stable and accurate, and it was more similar to the published values. But the procedure for further purification of DNA should be included.Conclusion The Light Cycler is approprate for clinical diagnosis of bacteria because. The HPLC method is accurate and reliable for bacterial classification.