Wash buffer that has had ethanol added should be clearly marked as such with a black X on the top, with a date of addition written.
For example, add 500µL of binding buffer to 100µL PCR sample.
CRITICALMake sure to label the silica columns.
This step ensures any residual ethanol, which may interfere with subsequent enzymatic reactions, is removed.
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This protocol describes the method used for silica-membrane-based column purification of up to 10µg PCR products (100bp to 10kb in size) using the _____ PCR Purification Kit. The purified DNA may subsequently be used in downstream assays, including sequencing, microarray analysis, ligation and transformation, restriction digestion, labeling, microinjection, PCR, and in vitro transcription.
DNA adsorbs to the silica membrane in the spin column in high-salt conditions and a pH ≤7.5 provided by the binding buffer (eg. buffer PB in a QIAquick kit). Impurities such as primers, unincorporated nucleotides, enzymes, mineral oil, salts, agarose, ethidium bromide and dyes are washed away by the ethanol-containing wash buffer (eg. buffer PE in a QIAquick kit). Pure DNA is eluted under low-salt and basic pH conditions with a small volume of elution buffer (eg. buffer EB in a QIAquick kit).