New! Clean-up problematic DNA in half the time of the original PowerClean® DNA Clean-Up Kit
The PowerClean® Pro DNA Clean-Up Kit utilizes our patented Inhibitor Removal Technology® (IRT) to provide researchers with a novel and proprietary method for cleaning up previously isolated genomic DNA. This kit is a significantly streamlined improvement over the original PowerClean® DNA Clean-Up Kit, with a 7 minute protocol that includes fewer steps. Starting DNA may be amber to brown in appearance; an indicator of PCR inhibiting substances, particularly humics and polyphenols. Even samples that appear colorless may contain PCR inhibitors which can be cleaned up with this kit. The PowerClean® Pro DNA Clean-Up Kit will remove this brown color as well as any PCR inhibiting substances, such as humic acids, heme, polysaccharides, polyphenols, fulvic acids, lipids and dyes. The resulting high purity DNA is ready to use in the most demanding downstream applications, including PCR, qPCR and next generation sequencing. This kit was validated with DNA isolated from a variety of problematic soils and also with DNA samples spiked with commercial humic acids. However, it performs well on DNA isolated from virtually any sample source.
Archived or previously isolated DNA samples are purified when combined with our proprietary DNA Clean-Up reagents. Inhibitors are selectively removed from the DNA solution. All DNA, including total genomic DNA, is captured on a silica membrane in a spin column format. DNA is then washed and eluted from the membrane. Percentage recovery varies depending on the level of inhibitors in the DNA that may be influencing the DNA yield measurement. Purified DNA is ready for PCR analysis and other downstream applications. We recommend quantifying DNA via a PicoGreen assay or running DNA on an agarose gel to determine recovery yield.
[Starting] [PowerClean® Pro] [Zymo OneStep™]
|PowerClean® Pro DNA||76.85||1.95||2.04|
[Starting] [PowerClean® Pro] [Zymo OneStep™]
Figure 1. Elimination of humic substances from DNA. Samples cleaned up with either the PowerClean® Pro DNA Clean-Up Kit or the Zymo Research OneStep™ PCR Inhibitor Removal Kit were examined on a 1.2% TAE gel (A). The amount, quality and molecular weight of DNA was similar for all samples, regardless of clean-up method. DNA quantification was performed using a NanoDrop 1000 Spectrophotometer (B). The concentration of the starting sample was observed to be artificially high, with low 260/280 and 260/230 ratios, indicating the presence of contaminants that absorb at A230, such as humic substances. Following clean-up with the PowerClean® Pro DNA Kit, the 260/280 and 260/230 ratios increased to levels consistent with pure DNA and the concentration of the DNA decreased to an average of 76.85 ng/μl, a value that was confirmed by quantitation using a Qubit PicoGreen Assay (data not shown). For samples cleaned-up with the OneStep™ Kit, the 260/280 and 260/230 ratios remained low, indicating that inhibitors were still present. Starting samples were brown due to the presence of humic substances (C). Following clean-up with the PowerClean® Pro DNA Kit, the samples were clear. However, samples processed with the OneStep™ Kit retained some brown color, indictaing that not all inhibitors were removed.
Figure 2. Successful qPCR amplification. DNA described in Fig. 1 was examined via qPCR with a Bacillus 16s assay (1 µl of undiluted DNA cleaned up using two different methods, or 1:10 dilution). Samples cleaned up with the PowerClean® Pro DNA Kit were free of inhibitors, as indicated by successful amplification and a difference of approximately 3 cycles between the undiluted DNA and the 1:10 dilution. Undiluted samples cleaned up with the Zymo OneStep™ Kit failed to amplify, and 1:10 dilutions amplified with a higher Cq value than the PowerClean® Pro 1:10 dilution, indicating that PCR inhibitors remained in the samples.
|Format||Silica Spin Filter Tubes|
|Method||Secondary DNA Clean-up|
|Binding Capacity||Up to 20 µg per filter|
|Sample Size||Up to 100 µl of purified DNA|
|Equipment Required||Vortex and Microcentrifuge|
Comparison with the PowerClean® DNA Clean-Up Kit
|PowerClean® DNA Clean-Up Kit||New PowerClean® Pro DNA Clean-Up Kit|
|Protocol Time||14 minutes||7 minutes|
Comparison with other Clean-Up Kits
|PowerClean® Pro DNA||GelSpin||Ultra-Clean® 15|
Silica Binding Particles
|Size Range||10bp - 50kb||60bp - 50kb||100bp - 50kb||
60bp - 50kb
|Solution or Reaction Clean-Up||X||X||X||X|
|Capacity||20 µg||20 µg||Scalable||20 µg|
|Inhibitors Removed||Environmental, salts, proteins||Salts, proteins||Salts, Proteins||Salts, Proteins|