A myo1Δgcn2Δ double mutant was constructed by replacing the MYO1 gene with a HIS5 module by homologous recombination using a PCR- based method.
SNP genotyping was determined using the ABI 7900HT Taqman SNP genotyping system (Applied Biosystems, Foster City, California, United States), which uses a PCR-based allelic discrimination assay in a 384-well plate format with a dual laser scanner.
A PCR based strategy using degenerate phr-specific primers was designed to detect and analyze possible photolyase genes.
One hundred ng of the DNA were used as a template in the genotyping analyses using a PCR-RFLP based method as reported by Engelke et al. [ 25].
The clonality of clinical and water M. abscessus isolates and a control strain (ATCC 19977) was determined using a rep-PCR based method (Diversilab® system, bioMerieux, Melbourne).
In an attempt to understand the mechanisms underlying this distinctive feature of TLT humoral response, an analysis of the TCR repertoire was performed using an RT-PCR based approach named "Immunoscope" [18].
These predicted exons were then tested for expression using an RT-PCR based strategy.
Using an RT-PCR based approach, we have shown that at least 8 different mutant transcripts are produced from the T Wis allele.
Before we used this PCR based test to detect M. fermentans, we confirmed the specificity of these primers.
CPV DNA titers were calculated by using a real-time PCR, based on TaqMan technology and able to recognize all CPV strains (10 ), whereas characterization of the viral type was obtained by means of MGB probe assays specific for types 2a/2b and 2b/2c (5 ).
Therefore, we did not attempt to use PCR based approaches as we feel the microarray analysis is more accurate and reproducible without the primer bias of PCR based approaches.
