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HER2/CEP17 ratio >2.2 was considered amplified (based on guidelines from the College of American Pathology CAPP)/American Society of Clinical Oncology ASCOO) 2007).
PIPSL locus including the coding sequence (CDS) and 1 Kb flanking regions (mainly untranslated regions, UTR) were PCR amplified based on primers designed by Oligo.
Patients with HER2 ratios, that is total gene/total centromere relations, of 1.5 or more were considered as amplified based on recent statistical approaches of our group [ 27] and included in this study.
Partial sequences for the HKGs that allowed the design of qPCR primers were obtained by either combining the SL-primer with reverse HKG primers previously used for Symbiodinium C3 [ 82], or amplified based on EST sequences for Symbiodinium B1 (Table 3).
"Someone has made that assumption initially and then it has been repeated and amplified based on no evidence whatsoever," he told Guardian Australia.
The signal was amplified based on the Ab2 AuPd DNA toward the catalytic precipitation of 4-choloro-1-naphthol (4-CN).
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Whereas all four cases were classified as HER2 amplified based on manual FISH, 3 of these 4 cases were scored as equivocal and one case as not-amplified based on automated FISH.
Amplified fragments based on a FHDCFV-containing motif primer (PERO1, PERO2) had much more frequently a strong homology (E value<10−5) to peroxidase protein sequences in PeroxiBase and NCBI after BLASTX analysis (18 out of 23) than the ones based on a VSCADI-containing motif primer (PERO4, PERO5, PERO6) (Table S2).
Direct sequencing of PCR amplified products based on the polymorphism of housekeeping genes has also become available in many diagnostic laboratories.
117 Solid resists are processed by deposition from solvent and most commonly consist of either positive-tone novolac chemistry or positive or negative-tone chemically amplified resists based on pendant group removal or epoxy chemistry, respectively.
To confirm that ERBB2 qRT-PCR data could accurately determine HER2 status, we developed a DNA-based HER2 copy number assay that showed a high correlation with ERBB2 gene expression that more accurately captured the HER2 amplified cases based on central confirmation (Supplemental Fig. 2).
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