Finnzymes’ Phire® Plant Direct PCR Kit is designed to perform PCR directly from plant leaves and seeds without prior DNA purification. Fresh plants, plant material stored at +4 Degrees C or frozen are all suitable templates for this kit, as well as plant material stored on commercially available cards such as Whatman 903® and FTA® Cards. A list of plants tested with this kit is available at www.finnzymes.com/directpcr.
The kit employs Phire® Hot Start DNA Polymerase, a specially engineered enzyme with a DNA-binding domain that enhances the processivity of the polymerase. Phire Hot Start DNA Polymerase also exhibits extremely high resistance to many PCR inhibitors found in plants.
The Phire Plant Direct PCR Kit contains reagents and tools for two alternative methods: direct and dilution protocols. A 0.5 mm Harris Uni-Core™ puncher and a compatible Harris Cutting Mat™ are included for convenient sample handling. Dilution Buffer is included for optional sample treatment before PCR (see ‘Dilution protocol’ in guidelines for sample handling). It can be used to treat larger/more difficult samples (e.g. more fibrous or latexcontaining samples), or when multiple PCR reactions are performed from a single sample. It is also a useful choice when longer DNA fragments (> 1 kb) are amplified. The kit includes control primers for amplification of a highly conserved region of chloroplast DNA.1 The kit is recommended for end-point PCR protocols.