HRP标记DNA探针的制备及杂交条件探讨.pdfVIP

第26 卷　第2 期 贵 阳 医 学 院 学 报 Vol.26 　No.2 2001 年4 月 　　　JOURNALOFGUIYANG EDICALCOLLEGE　　　 2001.4 HRP DNA 何　燕, 　单可人 ( , 　550004) [　] :(HRP)DNA , 。 : (PBQ)HRP , PEI HRP-PBQ-P EI , DNA, , HRP , 。 :HRP DNA , , 。 :DN A , , 。 [] ;;DNA [] Q8 13.4 　　[] A 　　[] 1000-2707 (200 1)02-0117-03 Investigation ofthe PreparationofDNAProbe Labeled withHRP andtheConditions ofHybridization HE Yan, SHA N Ke-ren (Mo lecular B iolog ica l K ey L abora tory of Guiy ang Med ica l College, Guiy ang 550004, China ) [Abstract] Objective:T o make DNA probe abe ed w ith H RP and discu ss t he conditions of hy- bridization reactions. ethods:H RP can be oxidized by PBQ to have an active site, and then con- densated w ith PEI to become H RP -PBQ -PEI comp ex .T he comp ex can connect DNA w ith g u- tara dehy de.T his characteristic can be used to abe the DNA probes with HRP .T he resu t of the hybridization reactions can be detected because the subst rates are co o red by the H RP .Results:We get t he DNA probes successf u y and have the better condit ions of hybridization reactions.Conclu- sion:T he DNA probes abe ed with HRP is a good method w ith hig her veracity and sensitivity . [Keywords] horseradish peroxidase (H RP );genetic marker ;D NA probes 　　DNA DNA ), 。1/10 1 mo /L [ 1] , N aHCO3 2.7 μ5 %P EI (), D NA , , 5 mmo /L PBS (pH 6.8)37 ℃ , 20 h, , 4 ℃。 [ 2] , 1.2 　　λDNA 5 mmo /L PBS [ 3] 。 (pH 6.8)20 μ, 20 μ H RP DNA , H RP DNA