Common use of Cases Clause in Contracts

Cases. At the Unit Molecular Diagnostics, Department of Pathol- ogy, Leiden University Medical Center, The Netherlands, 222 cases recorded as familial-CRC (fulfilling either Am- sterdam II criteria for HNPCC, Bethesda criteria, or being registered as late onset familial [three or more cases of CRC all diagnosed at age >50 years]) were registered between November 1999 and December 2002. These cases were analyzed following the medical ethical guide- lines described in the Code Proper Secondary Use of Human Tissue established by the Dutch Federation of Medical Sciences (▇▇▇.▇▇▇▇.▇▇/▇▇▇▇▇▇▇▇▇▇▇/▇▇▇▇▇▇- bruik/CodeProperSecondaryUseOfHumanTissue.pdf). The mean age of diagnosis of the 222 patients was 54 years. Appearance of tumor sites was distributed as fol- lows: coecum, 27; left colon, 15; colon transversum, 3; right colon, 38; sigmoid, 30; recto-sigmoid, 19; and rec- tum, 35. In 55 cases, the location was unspecified. One hundred and thirty-one cases showed a microsatellite stable phenotype, 88 cases had a microsatellite (MSI) instable (MSI-high, 71; MSI-low, 17) phenotype, and in three cases, the phenotype was unknown. As a control group, lymphocyte DNA of 156 healthy Dutch blood do- nors was used. Before analysis, MassEXTEND analysis of c.827A>C was validated with a standard control panel of 96 human genomic DNAs (BD Biosciences Clontech). Normal colon, carcinoma tissue was collected as 0.6- mm-diameter punches with a tissue microarrayer (Beecher Instruments, Inc., Sun Prairie, WI) based on evaluation of hematoxylin- and eosin-stained slides. Con- ventional microdissection (dissection with a needle of selected areas from a 10 μm hematoxylin-stained paraf- fin slide under microscopic examination with an inverted microscope.) was performed on the 22 adenomas of case 02031. Furthermore, flow sorting was carried out in three carcinomas containing <60% tumor cells (case 02031, 02395, and 01362) and one metastasis (02031).12 Genomic DNA was isolated from FFPE using a chelex extraction method as described by ▇▇ ▇▇▇▇ et al.13 MassEXTEND Genotyping of c.827A>C DNA samples isolated from normal tissue of the patient and control group were genotyped with the MassEXTEND assay (Sequenom, Inc., San Diego, CA). This SNP scoring is based on the mass difference of allele-specific primer ex- tension products. Design of the assay Figure 1. First, a 110-bp amplicon was generated using a standard PCR protocol with a forward primer, 5'-ACGTTGGATGGT- TCAATACAACATCAACCCG-3', and a reverse primer, 5'- ACGTTGGATGTTGTAACTCACCCAAGCCAC-3'. Note that

Cases. At the Unit Molecular Diagnostics, Department of Pathol- ogy, Leiden University Medical Center, The Netherlands, 222 cases recorded as familial-CRC (fulfilling either Am- sterdam II criteria for HNPCC, Bethesda criteria, or being registered as late onset familial [three or more cases of CRC all diagnosed at age >�50 years]) were registered between November 1999 and December 2002. These cases were analyzed following the medical ethical guide- lines described in the Code Proper Secondary Use of Human Tissue established by the Dutch Federation of Figure 1. Design of the MassEXTEND genotyping of c.827A�C SNP assay. 1) PCR amplification generated a product including the c.827A�C SNP. 2) MassEXTEND reaction that results in two products with different mass: c.827C allele, 6558.3 d, and c.827A allele, 7199.8 d. Medical Sciences (▇▇▇.▇▇▇▇.▇▇/▇▇▇▇▇▇▇▇▇▇▇/▇▇▇▇▇▇- bruik/CodeProperSecondaryUseOfHumanTissue.pdf). The mean age of diagnosis of the 222 patients was 54 years. Appearance of tumor sites was distributed as fol- lows: coecum, 27; left colon, 15; colon transversum, 3; right colon, 38; sigmoid, 30; recto-sigmoid, 19; and rec- tum, 35. In 55 cases, the location was unspecified. One hundred and thirty-one cases showed a microsatellite stable phenotype, 88 cases had a microsatellite (MSI) instable (MSI-high, 71; MSI-low, 17) phenotype, and in three cases, the phenotype was unknown. As a control group, lymphocyte DNA of 156 healthy Dutch blood do- nors was used. Before analysis, MassEXTEND analysis of c.827A>�C was validated with a standard control panel of 96 human genomic DNAs (BD Biosciences Clontech). Normal colon, carcinoma tissue was collected as 0.6- mm-diameter punches with a tissue microarrayer (Beecher Instruments, Inc., Sun Prairie, WI) based on evaluation of hematoxylin- and eosin-stained slides. Con- ventional microdissection (dissection with a needle of selected areas from a 10 μm �m hematoxylin-stained paraf- fin slide under microscopic examination with an inverted microscope.) was performed on the 22 adenomas of case 02031. Furthermore, flow sorting was carried out in three carcinomas containing <�60% tumor cells (case 02031, 02395, and 01362) and one metastasis (02031).12 Genomic DNA was isolated from FFPE using a chelex extraction method as described by ▇▇ ▇▇▇▇ De Jong et al.13 MassEXTEND Genotyping of c.827A>�C DNA samples isolated from normal tissue of the patient and control group were genotyped with the MassEXTEND assay (Sequenom, Inc., San Diego, CA). This SNP scoring is based on the mass difference of allele-specific primer ex- tension products. Design of the assay Figure 1. First, a 110-bp amplicon was generated using a standard PCR protocol with a forward primer, 5'-ACGTTGGATGGT- 5�-ACGTTGGATGGT- TCAATACAACATCAACCCG-3'�, and a reverse primer, 5'- 5�- ACGTTGGATGTTGTAACTCACCCAAGCCAC-3'�. Note that