Subsampling Ethanol Preservative from Zooplankton Museum Collections for DNA Extractions
Andreas Novotny
Disclaimer
Abstract
This protocol is used to sample DNA from archived samples preserved in ethanol, without having to subsample or split the actual zooplankton biomass.
Before start
Read background information, MIOP and BePOP-OBON information under the "Guidelines" tab.
Steps
PREPARATIONS
Prepare Sucrose Lysis Buffer (SLB):
Sucrose lysis buffer
ETHANOL SUBSAMPLING
To re suspend DNA in the zooplankton sample, invert the sample jar three times.
Let settle for 30 minutes
Use a serological pipette to remove 50 ml of the ethanol preservative, and transfer to a 50 mL falcon tube.
Use a syringe to push the ethanol through a
Seal the outflow of the filter with parafilm.
Add 1800 μL sucrose lysis buffer (SLB).
Seal the inflow opening with parafilm.
Label filter units and store them at -80°C for downstream DNA extraction.
DNA EXTRACTION
Follow the same extraction procedures as for Environmental DNA:
DNA Extraction from 0.22µm Sterivex Filters - Phenol-Chloroform
Alternative extraction method:
DNA Extraction from 0.22µm Sterivex Filters - Qiagen Blood and Tissue Kit
