The endothelial adhesion molecule, vascular adhesion protein-1 (VAP-1, AOC3) promotes lymphocyte recruitment to tumours, although the contribution that VAP-1 makes to lymphocyte recruitment in human colorectal cancer (CRC) is unknown. VAP-1 exists in circulating soluble form (sVAP-1). A previous study demonstrated elevated sVAP-1 levels in CRC patients. The aim of this study was to confirm this finding and study the differences in tissue VAP-1 expression between CRC and healthy tissues.

The MagNA Pure LC is an instrument for fully automated purification of nucleic acids (DNA, RNA and mRNA) from different sample types and volumes. The purification pricinple is based on the selective nucleic acid adsoprtion onto magnetic glass particles (MGPs), producing high quality nucleic acids. The instrument has walk-away processing and can perform 32 nucleic acid purifications in less than 60 minutes. The MagNA Pure LC requires no hands-on time after starting the protocol, and even the pipetting of subsequent PCRs can be done automatically. Therefore, the risk of contamination is significantly reduced whcih is crucial for all PCR analyses, and the reproducibility of hte results is improved. Moreover, due to the flexibility of the software, sample and elution volumes can be easily varied. The high quality of isolated nucleic acids allows you to perform numerous downstream applications such as PCR and RT-PCR analysis, enzymatic digestion, as well as Souther and Northern blot analysis.

High-resolution melting (HRM) allows single-nucleotide polymorphism (SNP) detection typing using inexpensive generic heteroduplex-detecting double-stranded DNA (dsDNA) binding dyes. Until recently HRM has been a post-PCR process. With the LightCycler 480 System, however, the entire mutation screening process, including post-PCR analysis, can be performed using a single instrument. HRM assays were developed to allow screening of the ryanodine receptor gene (RYR1) for potential mutations causing malignant hyperthermia (MH) and/or central core disease (CCD) using the LightCycler 480 System. The assays were validated using engineered plasmids and/or genomic DNA samples that are either homozygous wild type or heterozygous for one of three SNPs that lead to the RyR1 amino acid substitutions T4826I, H4833Y, and/or R4861H. The HRM analyses were conducted using two different heteroduplex-detecting dsDNA binding dyes: LightCycler 480 HRM dye and LCGreen Plus. Heterozygous samples for each of the HRM assays were readily distinguished from homozygous with both dyes. By using engineered plasmids, it was shown that even homozygous sequence variations can be identified by using either small amplicons or the addition of exogenous DNA after PCR. Thus, the LightCycler 480 System provides a novel, integrated, real-time PCR/HRM platform that allows high throughput, inexpensive SNP, and genotyping based on high-resolution amplicon melting.