DNA extraction - Zooplankton - 96 wells

Prepare your 96-well extraction plate with one individual per well. Alternate genus in the wells to detect eventual contamination between wells.

Collect one individual from a sample

Note its genus and determine its sex with a binocular microscope

For big individuals (more than 5 mm), dissect a few legs and put it in the well. Be careful not to damage the rest of the body and put it in a tagged Eppendorf tube.

For little individuals (less than 5 mm), put the whole body.

When all 96 wells are filled, the biological material has to dry to go to lysis

Prepare the lysis

Mix 18mL and 2.5mL in a Multi-Channel Reservoir and distribute 200µL with a multimicropipette in each well

Close your extraction plate with a heated aluminium foil and an adhesive plastic film

Put your plate in a proofer at 56°C to lyse the tissues

Perform the DNA extraction with a DNA extraction robot

Remove the adhesive film and aluminium foil from the plate and put it in the robot

Retrieve the new extraction plate containing the genomic DNA and discard the rest

It will deposit 200µL and 200µL in each wells and mix it

Then, 600µL are transfered on the tissue binding plate. Reagents excess are emptied in a waste container.

The tissue binding plate is then dried by a 0h 5m 0s aspiration to bind DNA to the silica membrane of the binding plate

The silica membrane is then washed with 600µL and twice with 900µL per well. Each wash is intercalated by a 0h 5m 0s aspiration dry.

The waste container is then removed from under the binding plate which is dried again by a 0h 10m 0s aspriration

An empty extraction plate is placed under the binding plate to retrieve the genomic DNA from it

DNA is eluted from the tissue binding plate with 100µL in each well and is collected in the new extraction plate underneath

After a 0h 3m 0s rest, the binding plate is dried for 0h 2m 0s and the elution is repeated with 100µL