QIAquick PCR Purification Kit

Many companies supply these kits, for example, the QIAquick PCR Purification Kit (Qiagen) or Montage PCR Centrifugal Filter Devices (Millipore). 

PCR purification kit or gel extraction kit (QIAquick PCR Purification Kit, QIAquick Gel Extraction Kit, both from Qiagen, or NucleoSpin Extract Kit from Macherey-Nagel)... 

QIAquick PCR Purification Kit (Qiagen) The features of the kit are available on the Web site http //www.qiagen.com/Products/ Catalog/Sample-Technologies/DNA-Sample-Technologies/ DNA-Cleanup/QIAquick-PCR-Purification-Kit. 

Two different commercial PCR purification methods are used. The ExoStar (GE Healthcare) method is a one-step enzymatic cleanup of PCR products that eliminates unincorporated primers and dNTPs. The QIAquick PCR Purification Kit (Qiagen) silica-membrane-based purification of PCR products (wNote 14). 

QIAquick PCR Purification Kit and the QIAquick Gel Extraction Kit (Qiagen, Valencia, CA). 

Purify the amplified DNA using the QIAquick PCR Purification Kit. Elute into 50 pL of the supplier-provided elution buffer. 

Split the purified cDNA from step 10 into two samples of 25 pL each. Mix each sample with 10 pL of the appropriate lOx reaction buffer (provided by the supplier) and bring to a total volume of 100 pL with water. Add 10 U of either of two restriction enzymes (Nla III or Tai 7 see Note 5) and incubate at the appropriate temperature (37 °C for Nla III, 65 °C for Tai I) for 1 h. Recover the digested cDNA using the QIAquick PCR Purification Kit, following the manufaaurer s protocol to the letter. Elute the cDNA in 50 pL of the included elution buffer (EB). 

Purify tlie PCRproducts using the QIAquick PCR Purification Kit, with a final elution volnme of 50 iL (r Note 11). Assess the resulting preparations nsing a Bioanalyzer and by agarose gel electrophoresis (Fig. 3a). 

Purify the cDNA using a QIAquick PCR Purification Kit as described in preceding sections. Elute the final cDNA sample into 50 pL. 

PCR PURIFICATION It is important to remove excess primers, nucleotides, enzymes, mineral oil, salts, and other impurities from DNA samples prior to executing other downstream processes. This DNA purification protocol is based on the QIAGEN QIAquick PCR Purification Kit (www.qiagen.com) however, several other DNA purification kits are available. We do not endorse this product but offer the outlined protocol as a starting place for the researcher. 

Purification of PCR products QIAquick PCR Purification Kit (Qiagen) or similar. 

Purify the PCR products with QIAquick PCR Purification Kit or similar. Follow the manufacturer s instructions and elute the product in the final step using 30 pL of ddH O. 

Purify the PCR products with QIAquick PCR Purification Kit following manufacturer s instructions. 

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