[Fluorescent mRNA differential display technique].
OBJECTIVE
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.


CONCLUSIONS
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.
Wang
G S
GS
Department of Gastroenterology, General Hospital of Chinese PLA, Beijing 100853.
Wang
M W
MW
You
W D
WD
Wang
H F
HF
Feng
M F
MF
D002460:Cell Line; D005453:Fluorescence; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D009000:Monocytes / Q000166:cytology