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目的 探索香港海鸥形菌(Laribacter hongkongensis,LH)DNA脉冲场凝胶电泳技术的条件。方法 从2株LH参考株中提取DNA,经过限制性内切酶SpeI的切割,再用脉冲场凝胶电泳仪进行DNA分子分型,比较5种条件的分型效果。结果 在OD值为1.8、细胞裂解4 h、SpeI 20U酶切20 h、电泳脉冲时间为2.2～54.2 s、电泳25 h的条件下,LH基因组DNA片段得到良好分离。结论 上述条件产生的LH DNA指纹图谱图像清晰,分辨率高,在没有图像分析仪的情况下,肉眼即可进行判断。证实所建方法可行,条件适合。
Objective To explore the conditions of DNA pulsed-field gel electrophoresis of Laribacter hongkongensis (LH) in Hong Kong. Methods DNA was extracted from two strains of LH reference strains, cut with restriction endonuclease SpeI, and then subjected to DNA molecular typing by pulsed-field gel electrophoresis. The typing effects of five conditions were compared. Results The genomic DNA fragments of LH were well separated at OD 1.8, cell lysis 4 h, SpeI 20U digestion 20 h, electrophoresis pulse time 2.2-54.2 s and electrophoresis 25 h. Conclusion The LH DNA fingerprints generated by the above conditions are clear and the resolution is high. In the absence of an image analyzer, the naked eye can be judged. Confirmed that the method is feasible, the conditions fit.