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PCR, qPCR, RT-PCR & dNTPs

Phusion® Human Specimen Direct PCR Kit

Product information

Features

• No need for time-consuming and expensive DNA purification steps
• Tested with a wide variety of sample types
• Two simple protocols for various applications
• Extremely short PCR protocol times
• Modified Thermo Scientific Phusion Hot Start II High-Fidelity DNA Polymerase guarantees high yields of specific product

Description

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 Phusion Human Specimen Direct PCR Kit kit is compatible with fresh, frozen, and even formalin-fixed, paraffin embedded human tissue samples.
The kit is based on modified Phusion® Hot Start II High-Fidelity DNA Polymerase. This specially engineered proofreading DNA polymerase is fast, robust and highly tolerant of many PCR inhibitors present in human tissues.
The Phusion Human Specimen Direct PCR Kit contains a complete set of optimized reagents and detailed instructions for various sample types. When combined with Piko® Thermal Cyclers and UTW® reaction vessels, PCR protocols can be completed in as little as 30 minutes. The kit is for research use only.

Content

Phusion® Human Specimen DNA Polymerase - 80 µl
2 x Phusion® Human Specimen PCR Buffer (includes dNTPs and MgCl₂) - 2 x 1 ml
Universal control primer mix (25 µM each) - 40 µl
Dilution Buffer - 2 x 5 ml
DNARelease™ Additive - 3 x 100 µl

Storage

Recommended storage temperature -20°C.

Choose direct or dilution protocol based on your needs

Two short and simple protocols for various applications. 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 0.5 μl 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.

Short protocols, superior performance

Fig. 1. Consistent results from various human samples with Direct PCR. 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. Reactions were run in UTW reaction vessels using Piko Thermal Cycler. + and – denote control reactions with and without purified DNA.
1. Buccal swab (0.5 mm punch)
2. Hair (1 mm)
3. Tooth (1 mm piece)
4. Nail (1x2 mm)
5. Saliva (0.5 µl)
6. Amniotic fluid (1.0 µl)

kb	Total cycling time (min)
	Direct PCR	Hot start Taq
0.5	30	84
1.0	30	84
1.5	35	99
3.8	62	202*

Fig. 2. Significant time savings and superior performance in comparison to conventional PCR from purified DNA. Four DNA fragments (0.5–3.8 kb) 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. The same DNA fragments were then amplified using a hot start Taq polymerase following the manufacturer's instructions. Phusion Human Specimen Direct PCR Kit delivered greater yields and was also able to reliably amplify the longer fragments. Comparison of total cycling times of both methods shows that the Phusion Human Specimen Direct PCR Kit is 2–3 times faster allowing significant time savings in PCR.

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Protocols & recommendations

RECOMMENDATIONS FOR USE