Genotyping For Trps1 Knockout Mice

Credit: Dr. Shivdasani, http://hms.harvard.edu I. Conditions for genotyping by genomic touch-down PCR: 94° – 30 sec 65° – 30 sec 5 cycles 72° - 30 sec 94° – 30 sec 60° – 30 sec 5 cycles 72°…

Megakaryocyte Culture From Mouse Fetal Livers

Credit: Dr. Shivdasani, http://hms.harvard.edu The protocol for culturing mouse megakaryocytes is essentially as described in the following publication: Lecine et al., Characterization of the transcription factor NF-E2 ..... J. Biol.…

Culture And Transfection Of Adherent Cells Fibroblasts Epithelial Cells

Credit: Dr. Shivdasani, http://hms.harvard.edu DMEM (5 °C/cold room) Dulbecco's Modidied Eagle Medium (500mL) 11995-065 Gibco L-glutamine (-20 °C) L-glutamine (200mM) 25030-081 Gibco Pen+step (-20 °C) Penicillin-Streptomycin…

Testing Rabbit Bleeds By Elisa

Credit: Dr. Mitchison, http://mitchison.med.harvard.edu Claire Walczak January 1996 You can test whether or not you have gotten an immune response to the peptide and how strong that immune response is by doing ELISAs against peptide conjugated…

Gene Transduction Of 1 Mks By Retroviral Infection

Credit: Dr. Shivdasani, http://hms.harvard.edu A. BSA (Bovine Serum Albumin) step gradient: Purify the megakaryocyte (MK) fraction from a day 2 liver culture (~10 livers/10 mL/10 cm tissue culture dish). 1) Pipet 1.5 mL of sterile 3%…

Genomic Pcr For Alleles Of Mouse P45 Nf E2

Credit: Dr. Shivdasani, http://hms.harvard.edu Prepare genomic DNA from tail biopsies, and resuspend in 150 μL sterile ddH2O. Heat DNA for 15-30 minutes at 65°C to dissolve. PCR Reaction: 10X Buffer 2.5 μL dNTP mix (5 mM each)…