The 3BPCR Clean-up kit offers a rapid and easy way to purify amplified DNA fragments directly from PCR reactions or from agarose gels (TAE/TBE). The technology behind this kit is based on membrane adsorption and isolation. This kit includes a binding buffer optimised for both kit uses. The kit ensures complete removal of salts, dNTPs, enzymes, primers, agarose, ethidium bromide and other impurities, while DNA fragments (65 bp – 10 kb) are bound to the silica membrane and purified with a high recovery rate. Support protocols included in the kit allow concentration and removal of salts, enzymes, etc. in samples containing SDS; and purification of single stranded DNA.
Procedure: DNA fragments from PCR reactions or agarose gels bind to the silica membrane-column in the presence of chaotropic salts from the binding buffer. Contaminants are removed by a simple washing step. The pure DNA is finally eluted under low ionic strength conditions with a slightly alkaline buffer or sterile bidistilled water.
For small double stranded DNA fragments, dilution of the binding buffer lowers the binding efficiency of the kit, without compromising the recovery of large PCR products. This particular feature of the binding buffer allows the exclusion of primer-dimers or side amplification products due to unspecific primer annealing. The obtained DNA fragments can be used directly in applications like automated fluorescent DNA sequencing, PCR or any kind of enzymatic reaction.
3B PCR Clean-Up kit components: Binding Buffer, Wash Buffer, DNA Binding Columns and Elution Buffer.
- Two in one kit with high recovery and small elution volumes (15 μl).
- Simple and easy procedure.
- Recovery of small DNA fragments <65 bp.
- Fast protocol: 10 min /6 preps.
- One buffer for both PCR clean-up and gel extraction
- Suitable for all polymerase buffer systems. <65 bp.
- Support protocol for cleaning up single stranded DNA.
- Support protocol for cleaning up samples containing SDS.