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3.5 反應(yīng)時(shí)間對(duì)體系熒光的影響 固定其它條件，考察BCDO-DNA體系在2、5、10、15、20、25、30(時(shí)間單位為min，數(shù)值從小到大，即由a→g)范圍內(nèi)7個(gè)不同時(shí)間對(duì)BCDO-DNA體系熒光猝滅的影響，結(jié)果如圖3.12和3.13所示。 根據(jù)圖3.12和3.13測(cè)定的熒光強(qiáng)度作BCDO-DNA體系熒光猝滅強(qiáng)度ΔF隨時(shí)間變化的曲線。結(jié)果如圖3.14所示。 由圖3.14可知，體系的熒光猝滅值ΔF大約在DNA加入5min之后才趨于平穩(wěn)。由此可知，小牛胸腺DNA與BCDO在六次甲基四胺-HCl緩沖溶液中大約5min反應(yīng)基本完成。所以依照2.2.4(1)的步驟，在加入DNA后，讓體系靜置大約5～10min左右使BCDO與DNA反應(yīng)基本完全后再進(jìn)行測(cè)定。 3.6 溶劑對(duì)體系熒光的影響 由于BCDO不溶于水，乙醇在這個(gè)體系中作為溶劑，所以需要固定BCDO和DNA的濃度，以及緩沖溶液的酸度，考察溶劑乙醇在2、5、10、15、20、25、30(單位為%，數(shù)值從小到大，即由a→g)范圍內(nèi)7個(gè)不同體積分?jǐn)?shù)對(duì)BCDO-DNA體系熒光猝滅的影響，結(jié)果如圖3.15和3.16所示。 根據(jù)圖3.15和3.16測(cè)定的熒光強(qiáng)度作BCDO-DNA體系熒光猝滅強(qiáng)度ΔF隨溶劑變化的曲線。結(jié)果如圖3.17所示。由圖3.17可知，當(dāng)乙醇的量保持在10%以下時(shí)，它對(duì)體系的影響較小，隨著乙醇用量的增加，對(duì)體系會(huì)產(chǎn)生較為嚴(yán)重的影響。 3.7 溶液溫度對(duì)體系熒光的影響 在實(shí)驗(yàn)中，固定其它條件考察溶液溫度(T)在20、25、30、35、40、45、50、60(溫度單位為℃，數(shù)值從小到大，即由a→h)范圍內(nèi)8個(gè)不同溫度對(duì)BCDO-DNA體系熒光猝滅的影響，結(jié)果如圖3.18和3.19所示。 根據(jù)圖3.18和3.19測(cè)定的熒光強(qiáng)度作BCDO-DNA體系熒光猝滅強(qiáng)度ΔF隨溫度(T)變化的曲線。結(jié)果如圖3.20所示，實(shí)驗(yàn)只需要在室溫條件下進(jìn)行測(cè)定。 |
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至尊木蟲 (職業(yè)作家)
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lina8804: 金幣+50, 翻譯EPI+1, ★★★很有幫助 2012-05-03 09:20:01
lina8804: 金幣+50, 翻譯EPI+1, ★★★很有幫助 2012-05-03 09:20:01
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3.5 The influence of reaction time to system fluorescence Fixing other conditions ,we inspected the fluorescence quench of system of BCDO-DNA in 2,5,10,15,20,25,30 minutes ,and the results figured in 3.12 and 3.13 . According to the figure of 3.12 and 3.13 ,we draw the fluorescence quench intensity with the changing of time ,which showed in figure 3.14 . According to the figure of 3.14 ,the system of fluorescence quench value would be balance after 5min of putting into DNA .We knew that the DNA of calf and BCDO would react completely after 5min of putting into HMTA-HCl .Therefore ,we let the system settled 5-10min after putting into DNA ,and then we detected the system . 3.6 The influence of solvent to the system fluorescence As a result of BCDO didn’t solve in water ,we set ethanol as solvent .We settled the concentration of BCDO and DNA ,and acidity of buffer solution ,then we inspected the fluorescence quench of system of BCDO-DNA in 2,5,10,15,20,25,30 % ,and the results figured in 3.15 and 3.16 . According to the figure of 3.15 and 3.16 ,we draw the fluorescence quench intensity with the changing of concentration ,which showed in figure 3.17 . According to the figure of 3.17 ,when the content of ethanol under 10% ,it had little influence in system ,but it had a serious effect with the increase of ethanol . 3.7 The influence of temperature to the system fluorescence Fixing other conditions ,we inspected the fluorescence quench of system of BCDO-DNA in 20,25,30,35,40,45,50,60℃ ,and the results figured in 3.18 and 3.19 . According to the figure of 3.18 and 3.19 ,we draw the fluorescence quench intensity with the changing of temperature ,which showed in figure 3.20 .The result just measured in room temperature . |
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