DNA and RNA Profiling in Human Blood: Methods and Protocols by Maryse St-Louis (auth.), Peter Bugert (eds.)

By Maryse St-Louis (auth.), Peter Bugert (eds.)

Blood samples have always confirmed to be a key resource of genetic fabric for a wide selection of diagnostic or learn reasons. In DNA and RNA Profiling in Human Blood: equipment and Protocols, prime overseas specialists give a contribution either confirmed and lately built protocols for advanced and high-throughput DNA and RNA profiling. Divided into thorough sections, the quantity concentrates on DNA profiling for blood mobile antigens via tools on high-throughput multiplex ways and SNP typing, besides RNA profiling in blood cells addressing sure blood telephone varieties corresponding to platelets, reticulocytes, and megakaryocytes. Written within the hugely profitable Methods in Molecular Biology™ sequence structure, the entire chapters contain short introductions at the topic, lists of the required fabrics and reagents, step by step, conveniently reproducible laboratory protocols, in addition to the Notes part which highlights pointers on troubleshooting and fending off recognized pitfalls.

Authoritative and state-of-the-art, DNA and RNA Profiling in Human Blood: tools and Protocols is a perfect advisor to the molecular profiling methods that experience spread out this vast box of analysis and feature proven nice promise within the additional selecting of illness markers in blood.

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9. TaqMan custom HPA assay probes, 15 μM (Applied Biosystems). 10. TaqMan 2x Universal PCR Mastermix with UNG AmpErase (Applied Biosystems). 11. Nuclease free water (Promega). Store at 4◦ C once open. 12. MicroAmp base/s (Applied Biosystems). 13. Pre-aliquoted DNA in MicroAmp 96-well optical plates. 14. DNA samples of known genotype homozygous for the ‘a’ and ‘b’ alleles of the HPA systems to be assayed at a concentration of 5–10 ng/μl. 3. Methods As with all PCR-based assays, particular care should be taken to avoid potential contamination of reagents and consumables at all times, ideally assay preparation should take place within a dedicated PCR set-up area or hood style workstation.

Transfusion 39, 763–771. 15. , Masera, G. (1985) Red cell alloantibodies in thalassemia major. Results of an Italian cooperative study. Transfusion 25, 110–112. 16. , Wagner, F. , Flegel, W. A. (2003) Antibodies to highfrequency antigens may decrease the quality of transfusion support: an observational study. Transfusion 43, 1563–1566. Chapter 3 Real-Time PCR Assays for High-Throughput Blood Group Genotyping Fernando Araujo Abstract There are multiple situations in the context of transfusion medicine where the classic serologic methods are unable to provide an adequate response, for example, recently polytransfused patients, patients with positive direct human antiglobulin tests, and hemolytic disease of the newborn.

1. Introduction A total of 24 serologically defined platelet alloantigens have been reported, 12 of which comprise the six well characterised and so called ‘common’ biallelic human platelet alloantigen (HPA) systems (HPA-1, -2, -3, -4, -5, and -15) (1). Although HPA-4 is considered a minor allele in Caucasoid populations. Twenty two of the 24 serologically defined HPAs have been identified at the molecular level, and with the exception of one system (HPA-14) all that differentiates between self and non-self in each case is a non-synonymous amino acid substitution resulting from a single nucleotide polymorphism (SNP) encoded in the relevant platelet glycoprotein (GP) gene (2).

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