71343  BloodDirect™ PCR Buffer Kit, Mouse

PCR amplification directly from mouse blood

The BloodDirect™ PCR Buffer Kits are novel buffers that neutralize DNA polymerase inhibitors present in mouse blood, eliminate all sample pretreatment steps, and allow direct PCR amplification from as little as 0.5 µl of blood treated with any commonly used anticoagulant.

Blood and other biological fluids contain a variety of Taq DNA polymerase inhibitors such as polysaccharides, proteins, lipids and their conjugates, hemoglobin, and heparin. Separation of these substances from the DNA template is necessary prior to PCR amplification. BloodDirect PCR buffers neutralize charge-bearing inhibitory substances that might bind to DNA polymerase and/or template DNA and ensure successful PCR amplification directly from blood.

Each BloodDirect PCR Buffer Kit includes two components: 5X BloodDirect Buffer 1 and either 5X BloodDirect Buffer A, in the human kit, or BloodDirect Buffer B, in the mouse kit. These buffers are added to the PCR mixture in place of 10X PCR buffer. The anticoagulant-treated blood sample (1 µl per 50-µl mixture, 0.5 µl per 20-µl mixture) is added as the final component.

BloodDirect Kits can be used in genotyping and genetic screening experiments with fresh blood samples, with samples stored at -20°C for up to four years, and with archived dried blood samples stored on filter paper or specimen collection cards. BloodDirect PCR Buffer Kit for mouse blood is ideal for routine transgenic mouse genotyping. The simple protocol dramatically reduces the risk of cross-contamination and sample mishandling.

BloodDirect Kits are compatible with commercially available Taq DNA polymerases, except for chemically-modified hot start DNA polymerases. To perform hot start PCR with BloodDirect PCR Buffer Kits, we recommend the use of any antibody-mediated hot start DNA polymerase, for example, NovaTaq™ DNA Polymerase plus Taq Antibody (Novagen).