HotSHOT DNA Extraction Kit

Rapid, affordable, and effective DNA extraction for PCR


HotSHOT DNA Extraction Kit

Rapid, affordable, and effective DNA extraction for PCR


Rapidly extract DNA from a wide range of samples with this widely used method. The HotSHOT DNA Extraction Kit is a simple, two step protocol performed in a single PCR tube. Provided as two 30 mL bottles containing an Alkaline Lysis Solution and a Neutralising Buffer, the kit includes enough reagents for up to 400 rxn.

2 x 30 mL


The HotSHOT DNA Extraction Kit provides users with a rapid, inexpensive, and reliable alkaline lysis method to extract PCR-quality DNA from a wide range of DNA-rich tissues. Suggested target tissues include animal tissues (from mammals to fish to invertebrates), although this protocol has also often been used successfully to extract fungal, plant, and bacterial DNA in a number of studies. It is a relatively crude DNA extraction method and is therefore not suitable for tissue types containing high proportions of PCR inhibitors.

This protocol was originally developed by Truett et al. (2000) for routine genotyping of mouse tissues. Since then it has been widely used in many different applications because of its low cost, simplicity, and scalability.

YouTube video


  • Versatile: Suitable for a wide variety of tissue types, typically from animal sources but also from plant, fungal and bacteria.
  • Simple: A two reagent process performed in one PCR tube per sample.
  • Scalable for low or high throughput: Can be used on a few samples or multiple 96-well plates. 
  • Rapid: Extraction of DNA in 30 minutes.
  • Affordable: Starting from 8¢ per sample, ideal for low cost approaches


Extraction of PCR-grade DNA from a wide range of sample types, for genotyping, PCR-based assays, sequencing, and DNA barcoding.

The HotSHOT alkaline lysis method is most often used for extracting DNA from mammal, bird, and fish tissues, but have also been used on a wide range of tissues from different taxonomic groups, ranging from invertebrates, animal parasites, and parasite blood meals; zooplanktonic eggs; pollen, and young leaves; and fungal pathogens in plant tissues.

The HotSHOT alkaline lysis method is not suitable for applications requiring extraction of large quantities of high quality unfragmented DNA. Electrophoresis gels of extracted genomic DNA are likely to be blank, or to show a faint smear of fragmented DNA and RNA, due to the small quantities of DNA extracted. However, this will not impact PCR results.


Recommended Usage

Add 75 µL of Alkaline Lysis Solution to 1-2 mm3 of a sample in a 0.2 mL PCR tube, and incubate at 95 ºC for 30 minutes. Allow to cool and then add 75 µL of Neutralising Buffer.

Use 1–5 µl of DNA extract as a template per 20 µl PCR reaction, or for difficult tissues (for example plant or fungal tissue, feathers, blood) first dilute 10x–100x in sterile distilled or PCR grade water to dilute PCR inhibitors. Optimal dilutions for a given tissue type can be established in the first instance by testing a range of dilutions.


Alkaline Lysis Solution (25 mM NaOH, pH 12)

Neutralising Buffer (100 mM Tris-HCl, 0.5 mM EDTA, pH 8)

Storage & Stability

Store sealed and in the original bag at room temperature. 

Exposure of the Alkaline Lysis Solution to air will gradually decrease the pH and effectiveness of this reagent over time. Storage for up to 6 months at room temperature should have minimal detrimental effects provided the containers and bags are properly sealed. 

For longer term storage, test the pH levels before use to check the pH of the Alkaline Lysis Solution.

Shipping conditions

Shipped at room temperature.

Related products

Adjustable Pipette

Agarose Tablets

100 bp DNA Ladder