A stringency wash was performed at 60°C for 30 min, and detection was performed using the 'In situ Hybridization Detection System' (K0601, DakoCytomation, Hamburg, Germany).
A stringency washes were conducted by using 0.5× SSC (Solution of Sodium Citrate) containing 0.1 % SDS (sodium dodecylsulphate), and the kit working solutions for appropriate time.
A stringency wash was then performed by rinsing slides in 2 × SCC, and the slides were placed in blocking buffer for 30 min in a humidified chamber at 37 °C.
Under the technical proposals, a stringency option of '9' suggested by the US would reduce overall aircraft emissions by 37.5%, while the '7' setting favoured by the EU would imply a 33% cut.
The raw 454 sequence reads from direct sequencing were imported into the CLC Genomics Workbench and trimmed at a stringency of 0.01 (equivalent to Q score of > 40).
Thermodynamic parameters obtained using the web server DINAMelt allowed supposing the possibility for realization (a) stringency control based on the ionic strength of the hybridization buffer and (b) stringency control based on the temperature elevation.
The raw sequences generated by HiSeq were imported into the CLC Genomics Workbench (Qiagen, Cambridge, MA), and trimmed at a stringency of 0.01 (equivalent to Q score of > 40).
Frequency analysis revealed that k-mer size was inversely proportional to the occurrence of k-mers in the different conserved regions, suggesting a stringency relationship; high numbers of duplicate reactions were observed with short k-mers, and a lower proportion of sequences were obtained with large ones, with the best results obtained using 12-mers.
This relation is the False Discovery Rate (FDR), a stringency indicator.
However, in all cases, the selection of genes undergoing expression variation is associated with a stringency parameter.
Applying a stringency gradient alone, resulted in 10 20% higher success rates (compare strategy 7 and 9).
