DNA extraction and purification kit Legionella

Next-generation DNA purification

→ Real-time PCR is a powerful and rapid tool for the specific detection of microorganisms in water.

This methodology, widely used in most countries, is already the subject of multiple ISO standards for the research of microorganisms, like Legionella spp & Legionella pneumophila, in various fields, such as environment or food industry.

Sample preparation (extraction and purification of nucleic acids) is the crucial step of the process.

With the DNA PURE-FLASH kit, our main goal was to eliminate PCR inhibitors while keeping optimal extraction performances.

→ The DNA PURE-FLASH  kit provides a robust and reliable solution for the optimal extraction of DNA from bacteria while ensuring the removal of inhibitors for real-time PCR analysis.

Application fields

The DNA PURE-FLASH kit allows an optimal DNA extraction of the microorganisms present in the water sample. It is used to prepare a sample in order to analyse it by real-time PCR. This methodology can be used on DRINKING WATER (treatment process, water supply units), SANITARY WATER (domestic hot and cold water systems, swimming pools, balneology…) or INDUSTRIAL WATER (cooling system…).

Protocol key points

1. Concentration of bacteria present in the sample by filtration or centrifugation, then, thermic lysis of the cells.

2. DNA retention using the Binding Buffer solution and creation of the polyplex.

3. Retention of the polyplex on a the WCX resin and elimination of the supernatant.

4. Addition of the elution solution and DNA recovery by orbital stirring at 80°C.

Advantages of
DNA PURE-FLASH technology

RAPID

extraction and purification in under 30 minutes

SIMPLE

clear and easy-to-use

EFFICIENT

elimination of PCR inhibitor

ROBUST

compatible with different matrix

PERFORMANCE

high extraction yields

FLEXIBLE

compatible with multiple PCR kits

RECOVERY

50%
on all matrix

VERY EFFICIENT PURIFICATION

less than 1%
of inhibited sample

SANITARY WATER
limit of detection

44 GU/l

INDUSTRIAL WATER
limit of detection

220 GU/l

Kit content for 96 reactions

– 96 tubes containing 1.3 ml of lysis buffer DNA PURE-FLASH,
– 4 dropper bottles (2.0 ml) of binding buffer DNA PURE-FLASH,
– 4 bottles (3.0 ml) of elution buffer DNA PURE-FLASH,
– 96 tubes of WCX resin,
– 96 microtubes of 2.0 ml,
– 96 microtubes of 1.5 ml,
– 192 tapered tips.

Equipment required

– Filtering manifold,
– Benchtop micro centrifuge,
– Polycarbonate membrane 0.45 μm,
– Thermomixer (temperature range: 25°C – 100°C / orbital stirring: ≥ 1500 rpm).

Optional:
– Vacuum pump,
– Ultrafiltration column.

We also recommend:

MIC qPCR cycler

For quantification of Legionella by qPCR.

Real time PCR kit for quantification of Legionella

DNA AMPLI-FLASH

Quantification kits of Legionella spp & pneumophila by qPCR.

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