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Thermo Scientific Phire Plant Direct PCR Kit is designed to amplify DNA directly from plant samples. No DNA purification is required prior to PCR. The kit is based on Phire Hot Start II DNA Polymerase, a specially engineered enzyme with a DNA binding domain. The unique features of this DNA polymerase make it extremely robust and tolerant of many PCR inhibitors present in plant material.
Phire Plant Direct PCR Kit has been tested with leaves and seeds from a wide variety of plant species. The kit includes a complete set of optimized reagents, sampling tools and detailed protocols making it an ideal choice for amplification of plant DNA.When combined with Piko Thermal Cycler and Piko UTW PCR Plates, PCR protocols can be completed in as little as 30 minutes.
In the direct protocol, a small piece of plant material is placed directly in the PCR reaction mix. In the dilution protocol, a small volume of the Dilution Buffer containing crushed plant material is added in the PCR reaction mix. The dilution protocol is used for long or difficult amplicons, and for assays where several DNA fragments are amplified from the same sample.
Various amplicons from mitochondrial or genomic DNA were succesfully amplified with Phire Plant Direct PCR Kit. 0.50 mm punches from different leaves were placed directly into 50 µL PCR reactions. Various amplicons from mitochondrial or genomic DNA were successfully amplified with the Phire Plant Direct PCR Kit. + and - denote control reactions with or without purified DNA.1 – Arabidopsis, mtDNA (1 kb)2 – Arabidopsis, gDNA (3.5 kb)3 – Capsicum, mtDNA (1.4 kb)4 – Maize, mtDNA (1.4 kb)
Punches (0.50 mm) from plant leaves or petal tissues were placed directly in 20 µL PCR reactions. 210 bp products were amplified using transgene specific primers. The results indicate that plant individuals corresponding to samples 2, 4, 7, 9, and 11 contain the RNAi transgene. The obtained results were confirmed by conventional analysis of CTAB DNA extraction followed by PCR. 1 through 9 – Leaves. 10 through 12 – Petals.