The L834R-EGFR pCDNA3 and T766M-EGFR pCDNA3 plasmids were produced through site-directed mutagenesis using the primers described in Supplementary Table 5.
The various loci were amplified by PCR using the primers described in Supplementary Table 5. Assays were performed with the Surveyor mutation detection kit (Transgenomics) as described41.
To test this possibility, we performed PCR using the primers described by these authors to amplify a region in the gadB gene of E. coli. Figure 6F shows that whereas the PCR was positive with a control sample of E. coli DNA, no fragment was amplified in the ten AD DNA samples.
Following the PCR reaction, 29 amplicons for faecal coliforms and 24 amplicons for Enterococcus spp. were sequenced using the conventional Sanger (dideoxy) sequencing in the forward direction using the primers described above.
Transgenic mice (line D) were screened by PCR using the primers described below.
The orientation of the cassettes in each clone was determined by PCR using the primers described in Table 1.
Then we carried out PCR experiments to amplify the entire ORF, using the primers described in Table S3.
Bissulfite-treated DNA was amplified by a nested-PCR protocol using the primers described in Table S2.
Genomic DNA spanning exon and nearby intron sequences was amplified using the primers described in Supplementary Table S8.
An RT-PCR using the primers described by Burmester and Plückthun was performed to amplify the VL and VH domains.
We amplified the coding sequence of the entire PYGM gene by polymerase chain reaction (PCR) in 14 fragments, using the primers described by Kubisch et al [21].
