Nucleic acid purification for use with PCR, quantitative real time PCR (qPCR) and the isothermal DNA amplification methods loop-mediated DNA amplification (LAMP) and recombinase polymerase amplification (RPA).
Pipette 100 μL of Extraction Buffer into a 1.5 mL tube. Add 1-2 mm3 of a sample to the tube. Grind the sample with a pestle, and dilute the sample with an additional 400 μL of Extraction Buffer. Dip a dipstick up and down 3 times into the extract, then dip 5 times into 1 mL of wash buffer. Dip the dipstick into 20–50 µl DNA amplification reaction mix 3–15 times (~10 s total) to elute.
Alternatively to store the sample, dip the dipstick into a tube of TE buffer. If necessary, molecular grade water can also be used for storage.
The dipstick purification method does not significantly concentrate the DNA sample, unlike solid-phase nucleic acid techniques. This means the method is best suited to PCR-based applications, and not suitable for methods such as restriction enzyme digests or clean up of PCR amplicons.
Due to the small capture volume of the dipstick, this method is unsuitable for purifying large quantities of nucleic acids, which means it is not suited to methods such as genome sequencing that require larger quantities of DNA For multiple PCR reactions from the same sample, an individual dipstick will be required.
Extraction Buffer (20 mM Tris-HCl, 25 mM NaCl, 2.5 mM EDTA, 0.05% SDS, 2% PVP-40, pH 8)
Wash Buffer (10 mM Tris-HCl, pH 8)
DNA Dipsticks (filterpaper, wax)
Storage & Stability
Store at room temperature (18–25 °C) for up to 1 year. For longer term storage, store at 4 ºC or freeze at –20 ºC.
Shipped at room temperature.