The GenElute™ PCR Clean-Up Kit is designed for rapid purification of single-stranded or double-stranded PCR amplification products (100 bp to 10 kb) from other components in the reactions, such as excess primers, nucleotides, DNA polymerase, oil and salts (Figure 1).
DNA is bound on a silica membrane within the spin column. The bound DNA is washed and the clean, concentrated DNA is eluted in the buffer of choice. Each column can purify up to 100 µl or 10 µg of PCR amplified DNA and recover up to 95% of PCR products between 100 bp and 10 kb. More than 99% of the primers and most primer-dimers (<40 bp) are removed.
Purified DNA can be used in enzymatic reactions, conventional or Automated Sequencing (Figure 2), and Cloning and Microarray Analysis.
Comparison of PCR product recovery and primer removal
Figure 1. Three separate PCR products were purified with GenElute PCR Clean-Up Kit and the kit from Supplier Q. Products were 143 bp from corn leaf, 375 bp from pBR322, 2 kb from human blood. Samples were analyzed on a 20% TBE acrylamide gel and visualized by staining with SyberGreen II™.
Figure 2. Sequence was resolved on an ABI 310 from a purified, 645 bp corn leaf PCR product. The PCR product was purified with the GenElute PCR Clean-Up Kit. The sequence was obtained by using ABI BigDye™ Terminator Chemistry and the same primers as for the original PCR.