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03-06-2014, 03:56 AM
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sequencing of pcr product cut from gel
Hello everyone! I am using QIAEX II Gel Extraction Kit to purify PCR products (300 bp) from agarose gel. The final elution is in 20 ul of H2O. I use 5 ul of the eluate in a BigDye Terminator sequencing reaction and then I purify my product using a Membrane-based filter plate. The sequence I get is unfortunately very bad quality , with some spikes and a lot of noise. Do you think I should change seq purifiction method?
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