PowerPrep™ Express PCR Purification Kits

Technology

The procedure involves dilution of the PCR reaction into binding buffer and loading onto a high capacity silica membrane to which the DNA is selectively adsorbed. After washing to remove enzymes, buffers, primers and dNTPs, the purified DNA is eluted in TE buffer or water and is ready to use in a variety of applications.

Performance Characteristics

Primers of up to 40 bp are completely removed with no loss of amplified products up to 12 kb (Fig 1 and 2). Longer primers may also be removed, but the efficiency is somewhat reduced (Fig 2). Removal of dNTPs, primers and DNA polymerase and column capacity and recovery are validated for each lot.

Fig. 1 Excess primer (40 bases) was added to PCR reactions containing a 1 kb DNA ladder. The ladder was then purified with the Marligen PowerPrep™ Express PCR Purification System. Lane 1 shows the unpurified mixture and Lane 2 shows the purified product.

Fig. 2 The effect of primer length on the efficiency of primer removal with the Marligen PowerPrep™ Express PCR Purification System.

It is sometimes desirable when purifying PCR fragments using 96-well plates to elute the DNA in a smaller volume so as to increase the final concentration of the solution. Figure 3 shows the effect of varying elution volume on the overall yield and the final concentration of the DNA recovered from the plate.

Fig. 3 The effect of different elution volumes on the DNA yield and concentration with the Marligen PowerPrep™ Express 96™ PCR Purification System.