Diagnostics and therapeutics for early-onset menopause
a technology for menopause and diagnosis, applied in the direction of instruments, drug compositions, immunological disorders, etc., can solve the problems of loss of overall bone mineral content, no diagnosis of hormonal analysis, and formidable challenges to the health care system, so as to effectively prescribe and treat or prevent the development of symptoms
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[0196] Determining an Individual's Allelic Pattern (Genotyping)
[0197] 1. IL-1RN (+2018); IL-1RN (VNTR); IL-1A (-889); IL-1B (+3954); IL-1B (-511); and IL-1A (+4845)
[0198] Sections 1.1 to 1.4 set forth general procedures that apply to Sections 1.5 to 1.10 unless otherwise indicated.
[0199] 1.1 Preparation of DNA. Blood is taken by venepuncture and stored uncoagulated at -20.degree. C. prior to DNA extraction. Ten milliliters of blood are added to 40 ml of hypotonic red blood cell (RBC) lysis solution (10 mM Tris, 0.32 Sucrose, 4 mM MgCl.sub.2, 1% Triton X-100) and mixed by inversion for 4 minutes at room temperature (RT). Samples are then centrifuged at 1300 g for 15 minutes, the supernatant aspirated and discarded, and another 30 ml of RBC lysis solution added to the cell pellet. Following centrifugation, the pellet is resuspended in 2 ml white blood cell (WBC) lysis solution (0.4 M Tris, 60 mM EDTA, 0.15 M NaCl, 10% SDS) and transferred into a fresh 15 ml polypropylene tube. Sodium ...
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