Sampling Tools for Direct PCR

Direct PCR master mixes for animal-friendly genotyping

PCR-based animal genotyping usually involves time-consuming DNA extraction steps from relatively large biopsies (1–5 millimeters) of ear, tail, or toe tissues prior amplification. Obtaining tissue samples of that size causes pain and distress to the animal, often requires anesthesia, and must adhere to animal-use regulation.

Invitrogen Platinum Direct PCR Universal Master Mix and Thermo Scientific Phire Tissue Direct PCR Master Mix allow for gene amplification from animal (including human) tissue samples without prior DNA purification (Figure 1). A sample is added directly to the master mix without the DNA extraction step, allowing significant savings in both time and cost. These master mixes allow robust PCR even in the presence of tissue-derived inhibitors, enables the use of short cycling times, and provides quick time-to-results. (Related: PCR tips for quick mouse genotyping)

Direct PCR. The direct PCR approach delivers convenience for DNA amplification by allowing PCR directly from samples without prior DNA purification

Figure 1. Direct PCR. The direct PCR approach delivers convenience for DNA amplification by allowing PCR directly from samples without prior DNA purification.

Due to their extraordinary sensitivity, the Platinum and Phire direct PCR master mixes require only small sample amounts for DNA amplification, which in turn minimizes the pain for the animal during the sample collection procedures. For tissue samples, a punch of 0.3–0.5 mm in diameter is sufficient for efficient PCR. In addition, the master mixes support PCR from animal hair—as little as a single hair follicle for robust DNA amplification (Figure 2).

Targets of different lengths were amplified from different number of mouse hair using Phire Tissue Direct PCR Master Mix

Figure 2. Targets of different lengths were amplified from different number of mouse hair using Phire Tissue Direct PCR Master Mix. 1: 237 bp, 2: 515 bp, 3: 1.1 kb, M: Thermo Scientific GeneRuler Express DNA Ladder.

Platinum Direct PCR Universal Master Mix and Phire Tissue Direct PCR Master Mix support animal-friendly, simple, and fast genotyping approaches. Don't you want to try genotyping the mice gently, just by using minimal samples?

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