Identification And Isolation Of Genes Differentially Expressed In Gastric Carcinoma By Optimised Differential Display Polymerase Chain Reaction

In this experiment, using cell line GC7901 and GES-1 as research targets, explored the optimised condition of mRNA DD-PCR, using optimised condition and GQS-960 type image and gene quantitive analysis system, cDNA fragments differentially expressed in GC7901 or GES-1 were isolated and sequenced as well as GenBank similar examination .Using obtained sequences ,designed 16 pairs of primers and applied them to examine clinical samples with ERK primers, aim was to establish gene diagnosis method for gastric cancer;and screened gastric carcinoma cDNA liabrary using GCYS-6 as probe , and did chrosome location by FISH and PRINSE.Results were as follow: optimised cycle parameters of mRNA DD-PCR were that : the first 5 round cycle parameters were that 94 ℃35s,40℃2min,72℃30s;and following 35 round cycle parameters were 94℃45s,55℃2min,72℃1min. One hundred and fifty four bands of cDNA fragments were obtained differentially expressed in cell line GC7901...