Items Tagged ‘Fero-Cell’

June 3rd, 2016

Purification of L-KS+ Marrow Cells

By UNM CC UNM CC

Purification of Lin-, c-kit+, Sca-1+ bone marrow cells for Culture, Flow Cytometry, or Transplantaion M. Fero 6/05 Materials Media Heat inactivated FBS (56°C x 30 min) PBS + 2% heat-inactivated FBS IMDM + 20% heat-inactivated FBS Viral transduction: Iscove’s + 20% heat inactivated FBS, plus 100 ng/mL each of SCF, IL-6, and Flt-3L, plus 10 […]

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June 3rd, 2016

Harvesting Hematopoietic Cells from Mice

By UNM CC UNM CC

M.Fero — May 5, 2003 Materials 4 mice from each genotype 4 Ly5 mice Buckets with wet ice 3x Bucket with dry ice 1x Dewar flask with liquid nitrogen 100 mL beakers with 95% ethanol 2x 100 mL beaker with dd. H2O 2x Sterile petri dishes Surgical tools Sterile IMDM medium aliquoted into 50 mL […]

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June 3rd, 2016

Simplified Calcium Phosphate Coprecipitation

By UNM CC UNM CC

M. Fero — 6/01 (after C Gorman, M. Calos) Materials 2x HEPES: 8 g NaCl, 0.105g NaHPO4, 6.5 g HEPES, H2O to 500 mL, pH 6.95 to 7.10 (try a range) 2M CaCl2: store in aliquots at -20°C DNA 4 mg/mL Procedure (The volumes can be scaled up as necessary) Let all solutions equilibrate to […]

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June 3rd, 2016

Culturing Mouse Embryonic Fibroblasts

By UNM CC UNM CC

(M. Fero 7.10.06) Materials Trypsin (Gibco 25200-023) 3T3 Medium: 500 mL DME (Invitrogen) + 50 mL FBS (Hyclone) + 5 mL 100x Pen/Strep 2x Freezing Medium: 3T3 Medium + 20% DMSO: Filter sterilized. Sterile PBS P100 and P60 tissue culture dishes Dissecting tools: Scissors, forceps, scalpel 95% ethanol in a squeeze bottle and in a […]

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