PureLink™ PCR Purification Kit, 50 preps - FAQs

View additional product information for PureLink™ PCR Purification Kit - FAQs (K310002, K310001)

6 product FAQs found

Can I use the PureLink PCR Purification Kit to clean up restriction digested plasmid DNA?

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

Can I use the PureLink PCR Purification Kit to do a gel purification?

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.

What are the minimum and maximum size ranges for DNA fragment cleanup using the PureLink PCR Clean-Up Kit?

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.

What is nested and semi-nested PCR?

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).

Can I sequence PCR products rather than plasmids, and what is the advantage?

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.

How can I purify my PCR product?

Life Technologies offers a number of PCR clean-up kits for standard and 96-well plate PCR. Search for PCR Clean-Up at www.thermofisher.com.