Mutational analyses in the three genes were brought by direct sequencing and each fragment was analyzed using an ABI 3730 DNA Sequencer.
The factor V Leiden mutation was determined by direct sequencing and the MTHFR genotype by a PCR-based RFLP method.
We analyzed the TERT promoter by direct sequencing and genotyping in 466 medulloblastomas.
Accession numbers of PRAT RNA monomers obtained by direct sequencing and cloning are EF209021-EF209033.
VHL mutation analysis in the VHL patient cohort was performed by direct sequencing and MLPA analysis and the mutation analysis results have been reported previously [20].
The ABL kinase domain of the BCR ABL allele was amplified with a PCR restriction fragment length polymorphism technique, followed by direct sequencing and sequence homology analysis.
We evaluated MASI status in seven mKRAS and two mEGFR lines using SNP arrays, and compared the results with MASI status determined by gene specific assays (mA% by direct sequencing and copy number by qPCR).
The aligned sequences, obtained by direct sequencing and cloning, were analyzed together with reference sequences from the HIV sequence database (http://hiv-web.lanl.gov/) using MEGA to exclude the possibility of contamination.
These and an additional set of 52 other genes that code for metabolic enzymes and transporters commonly involved in drug disposition were evaluated for the presence of known polymorphisms by direct sequencing and with a high-throughput SNPlex assay (Applied Biosystems) [32].
We determined 1) mutational status, 2) copy number gains (CNGs) and 3) relative ratio between mutant and wild type alleles of KRAS, BRAF, PIK3CA and EGFR genes by direct sequencing and quantitative PCR assay in over 400 human tumors, cell lines, and xenografts of lung, colorectal, and pancreatic cancers.
Genetic changes were confirmed by direct sequencing and correlated with biological and clinical data.
