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- W2354964480 abstract "Objective:To construct suppression subtractive libraries for identifying differentially expressed genes Methods:Total RNA was extracted from the whole brain tissue of normal and apoplectic stroke rats,from which mRNA was isolated to be the samples with different origination The experiment group was chosen as the tester and the control group as the driver The double-stranded cDNA synthesized by tester mRNA and driver mRNA was digested with restriction enzyme(RasI)to produce flush end Tester cDNA was divided into two parts to ligate two different types of adaptors while driver cDNA did not ligate adaptors Hybridization was performed twice between tester cDNA and driver cDNA Only the cDNA with different adaptors were amplified in the first polymerase chain reaction(PCR) Meanwhile,we performed a second subtraction in reverse Results:Total RNA typically exhibited two bright bands in ultraviolet rays,which corresponded to ribosome 28s and 18s RNA with the intensity ratio of about 1 5/1 The double-stranded cDNA appeared as a smear on sepharose electrophoresis after RsaI digestion,the average cDNA size became smaller After subtraction,the abundance of GAPDH decreased significantly Conclusion:The procedure represents an efficient means of constructing suppression subtraction libraries,which must be useful for identifying stroke-associated genes" @default.
- W2354964480 created "2016-06-24" @default.
- W2354964480 creator A5063684278 @default.
- W2354964480 date "2003-01-01" @default.
- W2354964480 modified "2023-09-25" @default.
- W2354964480 title "Construction of suppression subtractive libraries of brain tissue from apoplectic stroke rats and normal rats" @default.
- W2354964480 hasPublicationYear "2003" @default.
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