Sample preparation (for CARLA Kits)

To run our Cellular Activity RNA-based ElisA (CARLA), the “entry sample” consists of an RNA eluate originating from the water to be analysed. 

To prepare this material, 3 steps are necessary: (1) at the water sampling site – or in the lab, microorganisms are collected onto filters, (2) cells are then lysed to disrupt membranes and liberate their genetic content, (3) total RNA is chemically extracted and isolated.

  Volume of water / sample (for one taxonomic target) Filter porosity (polycarbonate or nylon) Transport and Storage
 Microalgae 1L 3-10µm1mL of RNA shield or lysis buffer
 Cyanobacteria 20mL 0.7-1µm

Filters in buffer can be kept at RT if processed rapidly, at +4°C for some days or frozen (-20°C) for long-term storage.

The Microbia Environment water sampler SOFiA has been designed for optimal and standardized on-the-spot filtration that simplifies storage and transportation of water samples. As a replacement, use a pump with the same characteristics.

We recommend to use a chemical cell lysis (incubation of filters in buffer at 60°C for 10 min).

Please, refer to the Microalgae and Cyanobacteria CARLA user manuals for detailed instructions.

For guaranteed final data interpretation, we recommend to use the Microbia Environnement Sample Preparation Kit (cat. #ME-SP-50) and the protocol adapted from the Quick-RNA Miniprep Kit from Zymo Research (ZR cat. #R1054).

Please, refer to the Microalgae and Cyanobacteria CARLA user manuals for detailed instructions.

Product name: Microbia Environnement Sample Preparation kit (50 prep.)

Product code: # ME-SP-50          Price: 300€ excl.tax. and shipping fees

Content:

For filter storage and cell lysis:

  • 2-mL tubes filled with Æ1-mm glass beads (x50)

For RNA extraction:

  • 800µl-capacity microcentrifuge filter columns (x50)
  • Wash buffer (24mL)
  • Lysis buffer (50mL)
  • Prep buffer (25mL)
  • 1.5-µL collection tubes

Please, refer to the Microalgae and Cyanobacteria CARLA user manuals for detailed instructions.

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