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||3-10µm||1mL of RNA shield or lysis buffer|
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.
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).
Product name: Microbia Environnement Sample Preparation kit (50 prep.)
Product code: # ME-SP-50 Price: 300€ excl.tax. and shipping fees
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