Recommended Usage

1. For each sample, pipette 200 μL of Extraction Buffer into a separate labelled 1.5 mL tube. Add 1-2 mm³ of sample (just enough to grind) to the relevant tube.
2. Grind the sample with a small plastic or glass rod pestle, then dilute the sample with an additional 300 μL of Extraction Buffer.
3. Dip the filter paper end of a dipstick (the white end) up and down into the extract 3 times to bind a small amount of DNA.
4. Add 1 mL of wash buffer into a new 1.5 mL centrifuge tube, and dip the filter paper end of the dipstick and handle into 1 mL of wash buffer 3–5 times.
5. Dip the dipstick into a 20 µl PCR reaction mix 3–15 times (~10 s total) to release the DNA.

Quantities of sample and Extraction Buffer can be adjusted to suit different types of sample:

• For samples rich in DNA and PCR inhibitors, use as little material as possible, and consider increasing the amount of Extraction Buffer.
• For extremely small samples, use less Extraction Buffer to increase the concentration of extracted DNA.

Read more about the process in our protocol for DNA Extraction using the Dipstick DNA Extraction Kit.

The dipstick method can be used for nucleic acid purification for use with PCR, quantitative real time PCR (qPCR), and isothermal DNA amplification methods such as loop-mediated DNA amplification (LAMP) and recombinase polymerase amplification (RPA).

The dipstick method has been used for tissue types including:

• Plant tissue (leaves of many species, including difficult plants like Citrus).
• Fungal tissue (macrofungi, microfungi, and lichens)
• Animal tissue (fluke eggs, crayfish, catfish, and salmon)

It’s a relatively recent method, and much of its potential is still to be explored!

Read here to find out more about whether the Dipstick DNA extraction method could suit your project needs.

Read about some uses of the Dipstick DNA Extraction Kit here:

• Leech et al. (2023). DNA-confirmed records of Cortinarius epipurrus and C. hirtus. Field Mycology, 24(3), 83-85. https://doi.org/10.63482/bg9k1e56
• Khajegi et al. (PREPRINT). Rapid Perioperative IDH1 Mutation Detection in High-Grade Gliomas using Novel LAMP assay. https://doi.org/10.21203/rs.3.rs-3207603/v1
• Ruszova et al. (2024). The Utilization of the SaLux19-Based Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid and Sensitive Identification of Minute Amounts of a Biological Specimen. Life, 14(5), 579. https://doi.org/10.3390/life14050579

Ambient shipping: This kit is shipped at room temperature.

No cold chain required: Because the reagents are chemically stable, there is no need for dry ice or insulated packaging, reducing shipping costs.

Unopened: Store at room temperature (18–25 °C) for up to 1 year.

Opened: Store at room temperature (18–25 °C) for up to 1 year. The Extraction Buffer and Wash Buffer can also be refrigerated (~4 ºC) or frozen (–20 ºC) to reduce the chance of microbial contamination or for longer-term storage. Please note that chilling or freezing can precipitate the detergent in the Extraction Buffer — to redissolve the detergent, place the Extraction Buffer bottle in a container of very hot water for 10–20 minutes until any precipitates are fully redissolved.

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: filter paper, wax, dye