For this purpose, semen from eight mature cocks were collected, initially evaluated (percent sperm motility, volume and concentration), pooled, assessed for motility, plasma membrane integrity, viability and acrosome integrity, and divided into six aliquots for dilution (1 5 °C7 °C) in Beltsville poultry, red fowl extender, Lake, EK, Tselutin poultry and chicken semen extenders.
Cells were cross-linked for 10 min with fresh formaldehyde (3 % final concentration), pooled as aliquots of 3 × 10 cells, and stored at −80 °C until use.
In one experiment, restimulation was also carried out by incubating the PBMCs with immortalized fibroblasts that had been loaded for 1 hour with 10 μM (final concentration) pooled 21-mer peptides overlapping by 11 amino acids and encompassing the entire sequence of the FIV-Pet Env.
Chloramine levels in pool water depend on chlorine and nitrogen concentrations, pool pH, temperature, and water circulation patterns (Hery et al. 1995; Massin et al. 1998; Nemery et al. 2002).
Due to lower concentrations Pools 1 4 were prepared for sequencing following the corresponding chapter in the TruSeq Custom Amplicon Library Preparation Guide, which uses heat denaturation instead of denaturation with 0.1 N NaOH.
The results with either concentration of dsPKCλ+ζ and dsCherry were equivalent, and therefore data from both concentrations were pooled together.
Fractions containing IN activity (eluted at 300 mM NaCl concentration) were pooled and concentrated or not by ultrafiltration (Centricon Millipore), followed by addition of 7 mM CHAPS.
Approximately 20 genotypes having, respectively, the highest and lowest NSC concentrations were pooled to constitute 2 contrasted 1-kg forage samples.
Additionally, the ratio of 95th percentile to mean in DLC distributions was found to vary by age, between dioxins, furans, and PCBs, and across mean, making a simple ratio approach impractical for describing population concentrations using pooled samples.
The amplification products were diluted to equal concentrations, then pooled in equimolar proportion and used as a substrate for the pyrosequencing reaction [24], [26].
