Various concentrations of MgCl2 and primers (primers on request) were used specific to each assay in a 10 µl final reaction volume.
The FRET calculator (Center for Live Cell Imaging, University of Michigan, available on request) was used to perform image processing based on the equations of FRET stoichiometry [28].
An ∼1000 bp PCR product from cDNA encoding the vaccinia virus G9 protein (sequence available on request) was used as dsDNA substrate.
Eight PCR primers and eight internal sequencing primers (sequences available from JT upon request) were used for bidirectional sequencing in subjects who carried homozygous genotypes or common alleles of interest.
A Perl script utilizing BioPerl modules (available upon request) was used for completeness analysis.
Primers flanking the targeted exons (sequence on request) were used to amplify the cDNA as described previously for dystrophin, but using a single PCR for 35 cycles.
An in-house program, MOTIFS (the program can be made available upon request), was used to identify motifs from the Multiple Sequence Alignment (MSA).
The interview guide (available from the corresponding author on request) was used flexibly so that the interviewer could explore patients' comments further if needed.
The resulting 38 codes (available on request) were used to analyze the final two transcripts for which interrater reliability was 0.82.
A newly developed real-time quantitative reverse transcription PCR (RT-qPCR) (primers and probes are available on request) was used to test additional samples from affected cattle farms.
