PureLink™ PCR Purification Kit

Yes, you can use our kit to clean up restriction digested plasmid DNA.

The buffers and procedure are different for the PureLink PCR Purification Kit and PureLink Gel Purification Kit. Therefore, you can swap the column, but the right buffer and protocol must be used.

The size range is between 100 bp and 12 kb. Our kit comes with two buffers, Binding Buffer (B2) and Binding Buffer HC (B3). The Binding Buffer HC (B3) eliminates primer-dimers and short failed PCR products that are smaller than 300 bp.

Nested PCR requires two separate amplifications, with the first one using one set of PCR primers and the second one using internal, "nested" primers and 1% or less of the first PCR reaction as a template. Nested PCR is used when the target is present in low abundance or when non-specific PCR products are being produced along with the specific product. Semi-nested PCR is used when there is only enough sequence information to make a primer internal to one end of the primary PCR product such as in RACE (rapid amplification of cDNA ends).

Yes, you can sequence PCR products. This is an excellent way to confirm the sequence of an insert when plasmid sequencing produces an unexpected sequence. The target DNA is amplified with a single set of primers and then sequenced using the same primers (although not all PCR primers work well as sequencing primers). Plasmid DNA can be PCR amplified using a one set of primers and the resulting product can then be purified using a column and then sequenced using the same primers. You can even get more specificity by using a sequencing primer that binds internally to one of the PCR primers.