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Phusion Blood Direct PCR Kit


PCR directly from whole blood without DNA purification.
  
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Blood IconThermo Scientific Phusion Blood Direct PCR Kit is designed for amplification of DNA from whole blood. It eliminates the need for a separate DNA purification step prior to PCR. The modified Phusion Hot Start II High-Fidelity DNA Polymerase is resistant to PCR inhibitors present in blood and retains its activity even with samples containing 40% whole blood.

This extremely accurate proofreading DNA polymerase makes the Phusion Blood Direct PCR Kit an ideal choice even for demanding applications, such as cloning. Phusion Blood Direct PCR Kit also allows analysis of blood samples with different anticoagulants, storage conditions and from various species.

Highlights

  • No need for time-consuming and expensive DNA purification steps
  • Simple protocol requires minimal hands-on time
  • Extremely short PCR protocol times
  • Modified Phusion Hot Start II High-Fidelity DNA Polymerase delivers abundant yields of specific product

The protocol is simple and requires minimal hands-on time

Includes

  • Phusion Blood II DNA Polymerase
  • 2X Phusion Blood PCR Buffer (includes dNTPs and MgCl2)
  • 50 mM EDTA
  • 50 mM MgCl2
  • DMSO
  • Universal Control Primer Mix

Examples of Materials Tested

  • Mouse blood
  • Pig blood
  • Cat blood
  • Dog blood
  • Cow blood
  • Bird blood
  • Human blood
  • Blood preserved with heparin, citrate, or EDTA
  • Blood stored on Whatman 903 and FTA cards
  
Storage Condition-20 C
HazardousNo
237bp fragments amplified from human, mouse, pig, cat, dog, cow, and bird blood.

Robust amplification from a wide variety of species

237bp fragments amplified from human, mouse, pig, cat, dog, cow, and bird blood.

Amplification of a 237 bp DNA fragment directly from whole blood of seven different vertebrates (5% blood in the reaction). + and - denote control reactions


Amplification of three DNA fragments from human blood collected on various filter papers. Piko® Thermal Cycler and UTW reaction vessels were used in PCR. + and - denote control reactions. <br /><strong>1</strong> – 903 Card<br /><strong>2</strong> – FTA Gene Card<br /><strong>3</strong> – FTA Elute Card

High yields from blood samples stored on various cards

Amplification of three DNA fragments from human blood collected on various filter papers. Piko® Thermal Cycler and UTW reaction vessels were used in PCR. + and - denote control reactions. <br /><strong>1</strong> – 903 Card<br /><strong>2</strong> – FTA Gene Card<br /><strong>3</strong> – FTA Elute Card

Amplification of three DNA fragments from human blood collected on various filter papers. Piko Thermal Cycler and UTW reaction vessels were used in PCR. + and - denote control reactions.

1 – 903 Card
2 – FTA Gene Card
3 – FTA Elute Card


Phusion Blood Direct PCR Kit amplifies from 1 to 40% blood in as little as 40 minutes. Supplier B amplified faint bands from 1-20% blood over 82 minutes. Supplier C's kit amplified bands from only 1 and 5% blood over 106 minutes.

Superior performance even at very high blood concentrations

Phusion Blood Direct PCR Kit amplifies from 1 to 40% blood in as little as 40 minutes. Supplier B amplified faint bands from 1-20% blood over 82 minutes. Supplier C's kit amplified bands from only 1 and 5% blood over 106 minutes.

The Phusion Blood Direct PCR Kit was compared to other kits designed for PCR directly from blood. A 588  bp genomic DNA fragment was amplified in the presence of increasing blood concentration in the reaction mixture. PCR was performed according to suppliers instructions using Piko Thermal Cycler and ultra-thin walled UTW reaction vessels. Total protocol times indicated at bottom. + and - denote control reactions.


Citations

  1. H-P. Fuehrer et al., Novel Nested Direct PCR Technique for Malaria Diagnosis Using Filter Paper Samples. J. Clin. Microbiol. 49, 1628-1630 (2011).

  2. N. Awad, A. E. El-Tarras, Genetic Analysis of Protein Phophatase 1 Regulatory Subunit 1 B (PPP1R1B) Gene among Autistic Patients. Aust. J. Basic & Appl. Sci. 5, 1-5 (2011).