For this purpose, chromosomal DNA was isolated by QIAamp DNA Blood Mini Kit (Qiagen) from the blood samples of rat offspring according to the kit procedures.
DNA extractions were carried out on all parents and offspring using DNeasy-96 kits (Qiagen, Valencia, CA) according to the manufacturers' protocol.
The expression of IgM, C3, TGF-β1, IL-1β, transferrin, lysozyme and their concentrations were detected respectively with quantitative real-time PCR and Elisa kits in offsprings at different stages from both naive and vaccinated brood stocks.
Beavers live in extended family units typically with a pair of adults, this year's kits, the previous year's offspring and sometimes older young.
Heterozygotes can be mated to produce offspring with two normal copies of Kit, but to the researchers' surprise, most of these progeny also had white patches, even though the mutant gene was no longer present.
DNA from parents and offspring was extracted with Qiagen DNeasy tissue kits.
DNA was extracted from the fin tissue of F0, F1, and 48 F2 offspring following manufacturer's recommendations (DNA easy kit; QIAGEN).
To determine the genetic status of each offspring, DNA was prepared using the DNeasy Tissue Kit (Qiagen Inc., Valencia, CA, USA) from a small tail snip taken between 2 and 4 weeks of age just prior to weaning.
Extraction and purification of total DNA from leaf tissues of the 195 sampled individuals together with their 3240 offspring was performed using a DNeasy 96 Plant Kit (Qiagen Inc., Hilden, Germany) as described in Fénart et al. (2007).
Offspring serum corticosterone levels were measured using a quantitative competitive ELISA kit (AssayMax Corticosterone ELISA Kit; AssayPro, St. Charles, MO, USA).
