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How to amplify PCR product - wonder why using linear fragment as template, PCR result was always sm (Dec/18/2007 )

Dear all,

I run PCR many times to amplify linear fragment[3 fragment assembled], but the result was so disappointed, most of the results: smeared, some meared but with the target thin bright band, when cut off the gel, PCR again, always smeared!!!

Can everyone give me some suggestions on PCR[the template is linear fragment], thanks, wish to get your reply sooner.



Without seeing your gel photo it is hard to say, but I know that if you load too much of your pcr product onto your gel you can get a smearing pattern with a brighter band corresponding to the correct product.
Try loading 1/4 of what you usually do.