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文献和实验(1000X) 0.5 ml + Glutamine (100X) 5 ml + 5% FBS (25 ml) ProcedureDraw venous blood into 60 ml syringe with 10ml 6% dextran + 7 ml citrate/citric acid. Fill to 60 ml total (blood 43 ml).Mix and sediment 30 min at RT (white blood cells now with serum
Preabsorbing Antisera with C. elegans Acetone Powder
graduated cylinder, and add 5 volumes of acetone. Mix by inverting several times; should see a large amount of floculent white precipitate. Allow the precipitate to sediment (takes ~30 min), and remove the supernatant by suction. 4. Wash the precipitate
ES / MEF cell culture and electroporation of targeting construct
: The MEFs should be 90% confluent (if they are not, feed them another day). Split as follows: Suction off old media, rinse flask x 1 with 10 ml phosphate buffered saline (PBS). Add 5 ml Trypsin/EDTA media (0.05%/0.02%) to the flask and incubate for 5 to 10
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