Title: DEVELOPMENT OF SILVER STAINING mRNA DIFFERENTIAL DISPLAY METHOD AND CLONING OF MORPHINE INDUCED GENES IN SY CELL
Abstract: Objective: To develop mRNA differential display polymerase chain reaction (DD PCR) method with silver staining, isolate and clone the morphine induced genes in SH SY5Y(SY) cell line. Methods: Total RNA was extracted from the SY cells treated with morphine. The cDNA copies of differential expressed mRNA species were amplified by DD PCR using an anchored primer 5'dT 12 MG, combined with two of arbitrary primers, respectively. The DNA bands on gel were displayed by silver stain method. After running on urea denaturing sequencing gel, the differential displayed DNA bands were visually recovered from the sequencing gel and reamplified by PCR. The reamplified products were cloned into pGEM T easy vector. Results: Several morphine induced genes were identified by the silver staining mRNA differential display established. Conclusion: The developed non radioactive differential display technique with silver staining was a simple, smart, and efficient method for isolating differentially expressed gene.
Publication Year: 1998
Publication Date: 1998-01-01
Language: en
Type: article
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