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- W1035485309 abstract "A prerequisite for studies of the expression of genes encoding connective tissue proteins throughout development is a procedure for the isolation of intact, biologically active messenger RNAs (mRNAs) from a variety of tissues. This chapter presents the procedure that facilitates the recovery of procollagen mRNAs from a calcified tissue—in this case chick embryo calvaria. Successful extraction requires the rapid disruption of the connective tissue accompanied by the instantaneous denaturation of tissue ribonucleases. This may be accomplished by violently homogenizing the frozen, powdered tissue in a potent denaturant, 5 M guanidinium thiocyanate, under strongly reducing conditions. The RNA is separated from the clarified homogenate on the basis of its buoyant density by sedimentation through a cushion of cesium chloride. The procedure allows tissue to be stockpiled in the frozen state. Because the RNA is initially recovered by sedimentation rather than a concentration-dependent precipitation very small quantities of tissue may be utilized. The chapter also describes a method for the initial identification of procollagen mRNA translation products after peptic digestion." @default.
- W1035485309 created "2016-06-24" @default.
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- W1035485309 date "1982-01-01" @default.
- W1035485309 modified "2023-09-28" @default.
- W1035485309 title "[8] Preparation and translation of interstitial (calvaria) procollagen mRNA" @default.
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- W1035485309 doi "https://doi.org/10.1016/0076-6879(82)82065-x" @default.
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