By Teresa V. Bowman, Akil A. Merchant (auth.), Meral Beksac (eds.)
Molecular equipment: Stem telephone Transplantation provides a compendium of state of the art study at the molecular steps enthusiastic about hematopoietic stem phone (HSC) activation and self-renewal. The emergence of HLA typing, and the confident impression it has had at the good fortune of medical transplantation is emphasised and mentioned via extraordinary stem telephone researchers.
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Additional info for Bone Marrow and Stem Cell Transplantation
As the Taq polymerase enzyme used in the PCR reaction does not have a 3Ј to 5Ј exonuclease proofreading ability, mismatched priming does not result in production of a PCR amplicon. In order for this approach to be successful and for PCR-SSP to be used routinely for HLA typing, it is necessary to optimise the annealing and other PCR conditions including buffer components, for each primer pair utilized. The amplicons produced, which are indicative of the presence of the target polymorphism within the sample DNA being tested, are visualized after agarose gel electrohoresis and staining, usually with ethidium bromide.
23. However, since the application of DNA-based methods, particularly after the introduction of the PCR, the sophistication of the methods available for HLA typing has increased significantly. This chapter will focus on the currently available DNA-based methods, their applicability for different tests and their advantages and disadvantages. 2. HLA Typing Methods The many HLA alleles found are the product of multiple single-nucleotide polymorphisms (SNPs) distributed singly or in groups “motifs” via recombinatorial mechanisms generating what is often referred to as a “patchwork” pattern of polymorphism.
Currently, the best success with transduction of human HSCs has been achieved using lentiviral systems. 26 Duda et al. The disadvantage to using retroviral transduction is that retroviral particles are only able to insert their DNA into the genomes of cells that are actively dividing. Protocols for retroviral labeling of bone marrow cells are thus carefully optimized for activation and stimulation of HSCs while preventing lineage differentiation. Lentiviral transduction systems do not require the target cells to be in cycle so are more appropriate for labeling cells that may not be actively dividing.