Homogenization of Fish Liver for Metabolomic & Transcriptomic Analysis

Sample: Stickleback fish (Gasterosteidae) liver

Protocol:

Add 50 - 150 mg (wet mass) stickleback fish liver to the salmon muscle to the 2 ml soft tissue homogenizing kit.
Add buffer: 8 ml/g methanol and 2.5 ml/g water*.
Run the Precellys at 6400 rpm, 2 x 10 sec with a 30 sec break.
For metabolomic analysis: extract with methanol, chloroform and water to separate the hydrophilic and hydrophobic metabolites. Store at -80°C until ¹H NMR spectroscopy.
For transcriptomic analysis: prepare total RNA using the Qiagen RNEasy DNA-free RNA kit.

*In this instance, the lab was using a buffer appropriate for the downstream isolation of both RNA and metabolites. You may substitute a more appropriate buffer for your application, if necessary.

Results:

The lab was able to carry out both omics techniques on the same liver homogenate.

Representative one-dimensional 1H NMR metabolite spectrum of stickleback liver prepared after homogenization in the Precellys 24.

Representative one-dimensional ¹H NMR metabolite spectrum of stickleback liver.

One of 48 mRNA expression subarrays from the stickleback 15k CDNA array prepared after homogenization in the Precellys 24

One of 48 mRNA expression subarrays from the stickleback 15k CDNA array.