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The reliable quantification of gene copy number variations is a precondition for future investigations regarding their functional relevance. To date, there is no generally accepted gold standard method for copy number quantification, and methods in current use have given inconsistent results in selected cohorts. In this study, we compare two methods for copy number quantification. beta-defensin...

The detection of minimal residual disease (MRD) using immunoglobulin and T-cell receptor (TCR) rearrangements as PCR targets provides important prognostic information on the in vivo effectiveness of treatment in acute lymphoblastic leukemia (ALL). Here we report on the real-time quantification of MRD in 25 ALL patients using LightCycler technology. We designed and adapted allele-specific oligon...

The detection of minimal residual disease (MRD) using immunoglobulin and T-cell receptor (TCR) rearrangements as PCR targets provides important prognostic information on the in vivo effectiveness of treatment in acute lymphoblastic leukemia (ALL). Here we report on the real-time quantification of MRD in 25 ALL patients using LightCycler technology. We designed and adapted allele-specific oligon...

Digital polymerase chain reaction (PCR) is a quantitative PCR technique with potential to be more accurate and precise than real-time quantitative PCR without the need for a standard curve. The digital PCR sample/reaction mix is randomly distributed into a large number of partitions, such that some partitions contain no copies of the nucleic acid template and others contain at least one copy. F...

Our objective was to use long PCR to determine the copy number of ptDNA and mtDNA in total DNA extracted from tissue samples. In previous work, the efficiency of amplification of DNA decreased as the target sequence length increased above about 1 kb, so that DNA quantification using long PCR was considered unreliable (Arezi et al. 2003). Here, we used a long-PCR endpoint procedure to amplify or...

Real-time PCR represents a new methodology that accurately quantifies nucleic acids. This has been made possible by the use of fluorogenic probes, which are presented in two forms, namely hydrolysis probes (also called TaqMan probes) and hybridisation probes. We decided to apply this methodology to cytokine mRNA quantification and this led us to the development of a protocol that provides an ea...

Quantification of human DNA has been an integral part of forensic DNA analysis. Hybridizationbased methods such as Quantiblot® kits were used extensively in the 1990s. These methods fulfilled the need at the time, since their sensitivity range was similar to the genotyping methods in use, such as restricted fragment length polymorphism. Later, the development of robust and more sensitive megapl...

Quantification of HIV-1 is important to quantify risk for disease progression as well as for acquiring infection associated with drug abuse. Prior quantification methods include immune and enzymatic procedures, e.g., quantifying HIV-1 p24 protein by ELISA and the Reverse Transcriptase by enzymatic assay. Improved quantification of HIV-1 RNA and cDNA was established using PCR. This paper describ...