hrmbdkc@hkucc.UUCP (04/26/89)
I WANT TO TRANSFECT CAT PLASMIDS INTO RABBBIT CHONDROCYTES. ---------------------------------------------------------- FEATURES OF CHONDROCYTES:- 1. CHANGE PHENOTYPE (DEDIFFERENTIATE) WHEN ALLOWED TO SIT DOWN ON CULTURE DISH. 2. FORM AGGREGATE WHEN ALLOWED TO GROW IN SUSPENSION. THE PREPARATION OF PRIMARY RABBIT CHONDROCYTES IS VERY TIME- CONESUMING. IT TAKES ONE WHOLE DAY TO DISSECT 3 RABBITS AND GET 180 MILLION CELLS. WHAT IS YOUR SUGGESTION OF CHOOSING WHICH TRANSFECTION METHOD THAT I SHOULD USE? AND PLEASE GIVE ME HINTS HOW TO DO THAT. CAT ASSAY USING SCINTILLATION METHOD ------------------------------------ I DOUBT THAT THE LEVEL OF EXPRESSION OF MY CAT PLASMIDS IN THE RABBIT CHONDROCYTES NOT HIGH. BY USING WHAT MODIFICATIONS CAN I MEASURE THIS LOW ACTIVITY. THERE ARE T7 AND T3 PROMOTER SEQUENCES IN MY CAT PLASMIDS. DO YOU THINK THIS WILL SEVERELY AFFECT THE ASSAY.