QIAquick PCR Purification Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
The amount of agarose excised from the gel should be as small as possible.	Remove the flow-through by aspiration. Avoid contamination of the collection tube rim.

Upstream tips
The amount of agarose excised from the gel should be as small as possible.
Protocol tips
Remove the flow-through by aspiration. Avoid contamination of the collection tube rim.

Publication protocol

Our initial experiments suggested that filter paper may function in commercial kits for nucleic acid purification. To investigate this possibility, we reassembled spin columns of Qiagen kits by replacing the original silica-based membranes with filter paper discs, then using the reassembled spin columns for purification of nucleic acids from various sources using respective Qiagen kits in our hands, including the DNeasy plant mini kit for plant genomic DNA, the RNeasy plant mini kit for plant RNA, the QIAquick PCR purification kit for PCR products, the QIAquick gel extraction kit for DNA extraction from agarose gels, and the QIAprep spin miniprep kit for plasmid DNA. For comparison of effectiveness, we also included original spin columns and reassembled spin column using glass fiber filters (Whatman glass microfiber filters, Grade GF/F, GE), a silica-based material usually adopted for recharging or preparing homemade spin column [9, 11].

To our surprise, the filter paper based-spin columns exhibited superior performance for purification of tomato genomic DNA and yielded higher final DNA quantities than the original Qiagen spin columns or the glass fiber-reassembled spin columns (Fig 2A). Filter paper-based spin columns were also able to purify plant total RNA (Fig 2B), DNA from PCR (Fig 2C), and to recover DNA from agarose gels (Fig 2D), although the yields of these nucleic acids were relatively lower compared to yields from the silica-based spin columns (Fig 2). Apart from these findings, we noticed that filter paper reassembled spin columns do not work well for purification of plasmid DNA using the QIAprep spin miniprep kit protocol, as indicated by weak plasmid DNA bands detected in agarose gel electrophoresis and lower concentrations measured using a UV spectrum meter (Fig 2E).

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Manufacturer protocol

Download the product protocol from Qiagen for QIAquick PCR Purification Kit below.

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