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Thermo Scientific Phusion Human Specimen Direct PCR Kit delivers unprecedented convenience for DNA amplification by allowing PCR directly from unpurified human samples. A tiny amount of sample is used directly in the PCR reaction with no prior purification steps. This allows significant savings in both time and cost.
Suitable sample materials include buccal swabs, saliva, amniotic fluid, hair, fingernails, teeth, and skin biopsies. The kit is compatible with fresh, frozen, and even Formalin-Fixed, Paraffin Embedded (FFPE) human tissue samples. It includes optimized reagents and detailed protocols for several sample materials. The modified Phusion Hot Start II High-Fidelity DNA Polymerase utilized in the kit is a highly accurate proofreading DNA polymerase; PCR products obtained are perfectly suitable even for cloning purposes. The kit is for research use only.
In the direct protocol, a tiny amount of solid or liquid sample is placed directly into the PCR reaction. In the dilution protocol, the sample is briefly incubated in incubation buffer and 0.5µL of the supernatant is used as a template in the PCR. The dilution protocol enables numerous PCR reactions to be performed from one tiny sample. For the quickest method from sample to results, choose the direct protocol.
†We recommend the dilution protocol and a longer incubation. Amplicon sizes less than 300bp are recommended due to fragmentation of DNA in FFPE samples.
Different samples were placed directly into 20 µL PCR reactions. A 237 bp DNA fragment was successfully amplified using the control primers included in the kit. + and - denote control reactions with or without purified DNA.
Four DNA fragments were amplified directly from buccal swabs using the dilution protocol. For the comparison reactions, DNA was first purified using a DNA purification kit from a major supplier followed by PCR with a hot start Taq DNA polymerase. Thermo Scientific Piko Thermal Cycler and UTW reaction vessels were used in PCR. Comparison of total cycling times shows that the Phusion Human Specimen Direct PCR Kit is 2 to 3 times faster allowing significant time savings in PCR.