A consequence of whole animal extractions and normalizing the libraries was the increased potential to sequence non-oyster transcripts, such as bacteria and algae.
Traditional methods of KGF1 production involve animal tissue extraction and prokaryotic expression in E. coli.
Cellulolytic enzymes are used in a large number of processes including supplementation of animal feeds, extraction of fruit and vegetable juices, pulp and paper manufacturing, starch processing, textile processing, and ethanol production [ 6– 10].
However, we were able to successfully amplify ribosomal 18S and mitochondrial cytochrome c oxidase subunit 1 (CO1) products from most of our tardigrade extractions – even from animals extracted from rehydrated herbarium samples that had been stored dry for 15 years.
M-EPC carried out the experiments involving MV of the living animals, mRNA extraction, and Northern blotting, performed histomorphometric countings, and drafted the manuscript.
We hypothesize that these subtle changes in expression could be due in part to the use of whole animal mRNA extractions for the screening of the arrays.
All qPCR experiments are made in duplicate and for some stages (D60, 82, 90 and adult), different animals, RNA extractions, and RT were used.
These issues are often mentioned in the academic literature and in SR fund prospectuses where they are used as negative screening criteria (e.g., child labor (social), animal experiments (moral), extraction of raw materials (ecological)).
Animal procedures (WAT extraction and MEF isolation) and study protocols were approved by the McGill University Animal Care Committee.
