AIM: To apply fluorescent mRNA differential display technique. METHODS: Total RNA samples were extracted from human monocyte line U937 treated/untreated with IFN and LPS, and were used as templates in differential display PCR. The anchored primers used were labeled with the fluorescent tag. After running on 5.6% denaturing PAGE gel, differentially expressed bands were excised and recovered, and finally reamplified. RESULTS: Three tested samples all showed amplified bands differed from 300 bp to 2.0 kb, the bands were bright and clear, the background was low. Both yes/no changes and upregulated/downregulated happenings were shown simultaneously. The reamplification bands were sharp and pure. CONCLUSION: We have successfully practiced fluorescent differential display technique in our lab. It is a fast, safe and cost-effective method used to sereen unknown expressed genes.
AIM: To apply fluorescent mRNA differential display technique. METHODS: Total RNA samples were extracted from human monocyte line U937 treated/untreated with IFN and LPS, and were used as templates in differential display PCR. The anchored primers used were labeled with the fluorescent tag. After running on 5.6% denaturing PAGE gel, differentially expressed bands were excised and recovered, and finally reamplified. RESULTS: Three tested samples all showed amplified bands differed from 300 bp to 2.0 kb, the bands were bright and clear, the background was low. Both yes/no changes and upregulated/downregulated happenings were shown simultaneously. The reamplification bands were sharp and pure. CONCLUSION: We have successfully practiced fluorescent differential display technique in our lab. It is a fast, safe and cost-effective method used to sereen unknown expressed genes.