Although the purification of DNA from agarose gels is routine, it is difficult and costly to purify large amounts of DNA using commercial gel extraction kits because of the limited capacity of the available columns.
Therefore, the development of affinity adsorbents for purification of DNA polymerases is of academic interest and practical importance.
After purification of DNA by Qiaquick DNA purification, DNA was quantified with a Light Cycler 480 (Roche) and the Light Cycler 480 release 1.5.1.62 SP software (Roche) using FastStart DNA essential DNA Green Master (Roche).
Both platforms are designed for parallel processing of up to 96 samples, and combine Qiagen's proprietary membrane chemistry with a vacuum manifold and pipetting robots to automate the purification of DNA and RNA from a variety of sample types.
Prior to the LAMP assay, a sample preparation protocol was applied that included a pre-enrichment step in buffered peptone water, followed by extraction and purification of DNA.
In the present study we describe the design, synthesis and evaluation of a combinatorial library of novel affinity ligands for the purification of DNA polymerases (Pols).
The method relies on a combination of positive and negative selection to eliminate time- and labor-intensive steps such as column cleanup and agarose gel purification of DNA during part assembly.
An efficient study design includes provisions for further processing of the original samples, such as cryopreservation of isolated cells, purification of DNA and RNA, and preparation of specimens for cytogenetic, immunological and biochemical analyses.
Purification of DNA fragments from gel or PCR mixtures were conducted with GeneJet PCR/Gel purification kits (ThermoScientific).
Purification of DNA was achieved using a Genomic DNA purification kit (Fermentas, GmbH, St. Leon-Rot, Germany) according to the manufacturer's instruction.
The PureYield™ Plasmid Miniprep System for isolation of E. coli plasmids and the Wizard® SV Gel & PCR Clean-UP kit for the purification of DNA fragments were obtained from Promega (Madison, WI, USA).
