【摘 要】
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A new method for rapidly detecting restriction enzyme pattes of Mycobacterium DNA using capillary electrophoresis with laser-induced fluorescence detection (CE-
【机 构】
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West China School of Public Health,Sichuan University,Sichuan,Chengdu Province 610041,China
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A new method for rapidly detecting restriction enzyme pattes of Mycobacterium DNA using capillary electrophoresis with laser-induced fluorescence detection (CE-LIFD)was developed.Polymerase chain reaction was used to amplify a 439-bp fragment of a 65,000-kDa(Mr)heat shock protein gene(hsp65)of Mycobacterium.After digesting amplification products by BstEII and HaeIII,pattes of enzyme cleavage products were detected by both CE-LIFD and agarose gel electrophoresis(AGE),respectively.Experimental parameters of CE were optimized.Restriction enzyme pattes of Mycobacterium DNA were detected in optimum electrophoresis conditions:a coated capillary column with a length of 50 cm and an inteal diameter of 100 μm,an electrophoresis buffer of 45 mmol/1 Tris-boric acid-ethylenediaminetetraacetic acid,and a running voltage of 11 kV.The restriction enzyme pattes for eight species of mycobacteria were studied.Relative standard deviations of the relative migration times of DNA segments were＜3.6%.Compared with AGE,CE is more outstanding in resolution and detection time,and it can be applied as a more effective means to DNA restriction enzyme patte analysis.
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