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Library construction was performed using the TruSeq Stranded mRNA library kit (Illumina, San Diego, CA).
Actual field construction was performed using TB asphalt mixtures with Sasobit® and using conventional asphalt mixtures.
Amplicon library construction was performed using 22 PCR fusion primers, containing genome target specific sequence with 454 sequencing adaptors.
Clustering and contig construction was performed using the TGICL software from TIGR [ 75].
Map construction was performed using a least squares approach implemented in the regression option in Joinmap.
The library construction was performed using restriction enzymes BsaAI and HincII following Hamilton et al. (1999).
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Phylogenetic analysis and dendrogram construction were performed, using MEGA5 software.
Target capture and library construction were performed using the protocol described in Hiatt et al.25 with modifications during library amplification.
Insert amplifications for plasmid construction were performed using High-Fidelity DNA polymerases (Pfu DNA polymerase [Promega] or the Phusion Hot Start II High-Fidelity DNA polymerase [Thermo Fisher Scientific, Waltham, MA, USA]) according to the manufacturer's instructions.
In ToxCreate the model construction is performed using IST web services, while the validation and reporting is executed using ALU-FR services.
Linkage analysis and genetic map construction were performed using JoinMap 4.0 software (http://www.kyazma.nl) [ 45].
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