DNAzol® Direct (DN 131)
DNAzol® Direct is a universal reagent for processing biological samples for direct PCR. No DNA isolation is required. The DNAzol Direct procedure is simple and fast. Lyse a sample in DNAzol Direct for 15 min, add an aliquot of the lysate to a PCR mix, and perform amplification of a selected DNA fragment(s). DNAzol Direct simultaneously acts to release and denature DNA into a single-stranded form, hydrolyze RNA, and denature and partially hydrolyze proteins. The patent-pending DNAzol Direct composition and procedure are based on an alkaline solution containing polyethylene glycol and other additives. The combined effects of the alkaline pH and chaotropic properties of this reagent are sufficient to effectively inactivate PCR inhibitors including proteases and nucleases. This protocol eliminates columns, centrifugation steps and DNA precipitation. The standard DNAzol Direct procedure supports PCR amplification of DNA fragments up to 8 kb long.
DNAzol® Direct is designed to process a wide range of samples including animal, plant, fungi, yeast, bacterial, and viral samples. It has been used with blood, serum and plasma, saliva, buccal swabs, blood cards, hairs and feathers, leaves and seeds, and formalin-fixed tissues.
- Mix 1-10 μI fluid or 1-10 mg of solid sample with 0.1 ml of DNAzol® Direct.
- Lyse the sample by incubation for 15 min at room temperature.
- Vortex the lysate and transfer 2-5 μI of lysate directly into a 20-50 μI PCR mix. The lysate volume should not exceed 10% of the reaction volume.