By Teresa V. Bowman, Akil A. Merchant (auth.), Meral Beksac (eds.)
Molecular tools: Stem phone Transplantation offers a compendium of state of the art learn at the molecular steps focused on hematopoietic stem cellphone (HSC) activation and self-renewal. The emergence of HLA typing, and the confident effect it has had at the good fortune of medical transplantation is emphasised and mentioned via awesome stem cellphone researchers.
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Extra resources for Bone Marrow and Stem Cell Transplantation
16000044) (500 mL). 11. Dulbecco’s modified Eagle’s medium (DMEM; Invitrogen, cat. no. 10569-010) (500 mL), or StemSpan SFEM (Stem Cell Technologies, cat. no. 09650) (500 mL). 12. Hexadimethrine bromide (polybrene; Aldrich, cat. no. 107689-100G). Stock concentration 5 mg/mL in PBS, store at 4°C. 13. HEPES Buffer Solution (1 M; Invitrogen, cat. no. 15630-080) (100 mL). 2 Lentiviral Transduction of Human HSCs and Progenitor Cells 1. Highly transfectable cell line to be used as packaging cells. This protocol describes conditions optimized for 293T cells.
The reverse PCR-SSO methods involve immobilization of the oligonucleotide probes on a solid-phase support, and subsequent hybridization of the solid-phase immobilized oligonucleotide probes with liquid-phase PCR product. Solid-phase supports most widely in use include: 1. Nitrocellulose membrane strips (Reli™ SSO Dynal Biotech, The Wirral, UK and INNOLiPA line probe assay, Innogenetics, Gent, Belgium). 2. Microtest plates (enzyme-linked probe hybridization assay [ELPHA], Biotest, Germany). 3. , Los Angeles, CA).
Immunol. 62, 850–857. 7. , et al. (2004) Polymorphism analysis within the HLA-A locus by universal oligonucleotide array. Hum. Mutat. 24, 428–434. 8. Li, A. , Yang, J. , and Iwaki, Y. (2004) Multiplexed analysis of polymorphisms in the HLA gene complex using bead array chips. Tissue Antigens 63, 518–528. 9. Olerup, O. and Zetterquist, H. (1992) HLA-DR typing by PCR amplification with sequence-specific primers (PCR-SSP) in 2 hours: an alternative to serological DR typing in clinical practice including donor-recipient matching in cadaveric transplantation.