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文献和实验Revised protocol for establishing a mammary cell line from tumors
Revised protocol for establishing a mammary cell line from tumors Drench mouse in 70% ethanol, pin and dissect open Sterile dissection of tumor- leave some behind to use in histology Place tumor in small culture dish Weigh
Culturing Mouse Embryonic Fibroblasts
on the 3T3 protocol will maximize the growth prior to the development of cellular senescence. The 3T3 protocol was originally described as the passage of 3 x 105 cells every 3 days on 50 mm. dishes which is equivalent to 1.2 x 106 cells on a P100 dish
Methylated CpG Island Amplification
Prepare tubes containing 10 m l of 10 X PCR buffer, 100 pmol of RXMA24 (or RMCA24) primers, 15 Units of Taq DNA polymerase, 1.2 m l dNTP mix (25mM), 0 m l (RXMA) or 5 m l (RMCA) DMSO, H2 O to a total volume of 97 m l. Add 3 m l of ligation mixture
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