363 qPCR䈮ᦨㆡ䈭✛⦡Ⱟశ⦡⚛ qPCRဇEvaGreenTM EvaGreenTM 䈲ቯ㊂⊛real-time PCR䋨qPCR䋩䈮䉋䉍ㆡ䈚䈢✛⦡Ⱟశ⦡⚛䈪䈜䇯 ․㐳 䃂㩷㪧㪚㪩ᔕ䉕䈾䈫䉖䈬㒖ኂ䈚䈭䈇 䃂㩷㕖Ᏹ䈮ᒝ䈇Ⱟశᒝᐲ 䃂㩷ථ䈚䈢ቯᕈ 䃂㩷ఝ䉏䈢ㆡวᕈ 䂓㩷ⰯశᒝᐲᲧセ Relative Fluorescence 7000 EvaGreen: A␠ຠ:ٌ 6000 5000 4000 3000 105 104 103 30 35 102 101 2000 1000 0 0 5 10 15 20 25 40 45 50 Cycle Number ࿑1. EvaGreenTM䋨྾ⷺ䋩䈫᳢↪A␠ຠ䋨ਃⷺ䋩䉕↪䈇䈢䇮 䊍䊃ᨴ⦡DNAቯ㊂䈮䈍䈔䉎PCR䈱േ⊛䈭䊒䊨䉾䊃䈪䈜䇯᳢ ↪A␠ຠ䉕↪䈇䈢DNAჇ䈲䇮⥋ㆡỚᐲ䈪ⴕ䈇䉁䈚䈢䇯 EvaGreenTM䈱Ⱟశᒝᐲ䈲᳢↪䈘䉏䈩䈇䉎A␠ຠ䈱Ⱟశ 䉋䉍䉅ᢙᒝ䈒䈭䉍䉁䈜䇯 䂓 PCR䈻䈱ᓇ㗀 ᳢↪A␠ຠ䈫䈲ኻᾖ⊛䈮EvaGreenTM䈲PCRᔕ䉕䈾䈫䉖䈬㒖ኂ䈚䉁䈞䉖䋨࿑2䈍䉋䈶࿑3䋩䇯 EvaGreenTM Gives Cleaner, Sharper and Stronger Melting Peaks EvaGreenTM Shows Little Inhibition to PCR 9000 A␠ຠ 1x A␠ຠ 0.5x EvaGreen 1x EvaGreen 0.5x 8000 7000 6000 Relative Fluorescence Change-d(RFU)/dT Relative Fluorescence ↪ 5000 4000 3000 2000 1000 EvaGreenTM A␠ຠ extra peak 0 0 10 20 30 40 50 Cycle number 80 85 Temperature (oC) 90 80 85 90 Temperature (oC) ࿑2. EvaGreenTM䈫᳢↪A␠ຠ䉕䈠䉏䈡䉏2⒳㘃 ࿑3. 4䈧䈱⇣䈭䉎PCR↥‛䈮䈧䈇䈩EvaGreenTM䈫 䈱⇣䈭䉎Ớᐲ䈪↪䈇䈢PCRჇ䇯᳢↪A␠ຠ䈲1 ᳢↪A␠ຠ䉕↪䈇䈢႐ว䈱DNA䊜䊦䊁䉞䊮䉫ᦛ✢ 䈱Ớᐲ䋨OD䌾0.05䋩䈪ᗧ䈭PCR㒖ኂ䉕䈖䈜 ⸃ᨆ䇯TBP䋨䂾䋩䇮SDHA䋨䂦䋩䇮RPL4䋨䂺䋩䇮 HMBS䋨䂔䋩䇯 TM 䈱䈮ኻ䈚䇮EvaGreen 䈪䈲㒖ኂ䈲䉌䉏䉁䈞䉖䋨ᝌ ᳢↪A␠ຠ䈏PCR䉕ὑ䈮㒖ኂ䈜䉎⚿ᨐ䇮ౣᕈ ࿑䉕ෳᾖ䋩䇯 䈱䈭䈇ಽ䈭Ⲣ⸃䊏䊷䉪䈱ᒻᚑ䈏䉌䉏䉁䈜䇯 ຠ⺑ 䂓㩷ቯᕈ 䈘䉌䈮䇮ਇቯ䈫ႎ๔䈘䉏䈩䈇䉎᳢↪A␠ຠ䈫䈲⇣䈭䉍䇮EvaGreenTM䈲䉝䊦䉦䊥ᕈ᧦ઙਅ䈅䉎䈇䈲 ㉄ᕈ᧦ઙਅ䈪䉅䇮ᾲ䉇ട᳓ಽ⸃䈮ኻ䈚䈩㕖Ᏹ䈮ቯ䈚䈩䈇䉁䈜䋨࿑4䋩䇯 Relative Fluorescence EvaGreenTM is Highly Stable 120% 100% 80% 60% 40% 20% ࿑㪋㪅㩷㩷 㪐㪍㷄䈪䈱㪛㪥㪘ሽਅ䈮䈍䈔䉎㪜㫍㪸㪞㫉㪼㪼㫅㪫㪤䈱ቯᕈ 0% 0 5 10 15 20 25 30 35 40 Time (min.) at 96oC 䉕␜䈚䈩䈇䉁䈜䇯⋧ኻᲧセ䈜䉎Ⱟశ୯䈲㪍㪇㷄䈪᷹ቯ䈘䉏䇮 ᤨ㑆䈲㪐㪍㷄䈮⛽ᜬ䈚䈢䊃䊷䉺䊦ᤨ㑆䉕␜䈚䈩䈇䉁䈜䇯 䂓㩷ๆ䉴䊕䉪䊃䊦 Emission Excitation EvaGreenTM䈱ๆ䈍䉋䈶⊒శ䉴䊕䉪䊃䊦䈲 ᳢↪A␠ຠ䈅䉎䈇䈲FAM䈮㘃ૃ䈚䈩䈇䉎䈱䈪䇮 EvaGreenTM䈱Ⱟశ᷹ቯ䈮䈲䇮ห䈛శቇ᷹ቯ䈱⸳ቯ䉇ⵝ⟎䉕↪䈜䉎䈖䈫䈏᧪䉁䈜䋨࿑5䋩䇯 400 450 500 550 ଔᩰ 䈱ബ䈍䉋䈶Ⱟశ⊒శ䉴䊕䉪䊃䊦䉕␜䈚䈩䈇䉁䈜䇯㕍⦡䈱 ✢䈲ബ䉴䊕䉪䊃䊦䇮✛⦡䈱✢䈲Ⱟశ⊒శ䉴䊕䉪䊃䊦䉕␜ 600 Wavelength (nm) ෳ⠨ᢥ₂ ࿑5. PBSਛ䈮䈍䈔䉎ੑ㊀㎮DNAሽਅ䈪䈱EvaGreenTM 䈚䈩䈇䉁䈜䇯 Karsai et al. BioTechniques 32(4), 790 (2002) ຠฬ ⾂ᴺ ࡔࠞຠ⇟ภ ኈ㊂ ଔᩰ EvaGreenTM, 20 -20͠ 31000 51 ml 34,100 EvaGreenTM qPCR BasicMix 2* -20͠ 31001 31.7 ml 34,100 *Basic Mix 䈮䈲㩷qPCR 䉕ⴕ䈉䈱䈮ᔅⷐ䈭䉅䈱ో䈩㩷(Taq㉂⚛䉕㒰䈒) 䈏䉁䉏䈩䈇䉁䈜䇯 ߏᵈᗧ˴⹜㛎⎇ⓥ↪એᄖߦߪߏ↪ߒߥߢਅߐޕ ̪ଔᩰߦߪᶖ⾌⒢ߪ߹ࠇߡ߅ࠅ߹ߖࠎޕ ⽼عᄁขᛒᐫ ࠽ࠞࠗ࠹ࠬࠢᩣᑼળ␠ 㩷 ˴ޥ੩ㇺᏒਛ੩ੑ᧦ㅢὖਣ᧲₹ደ↸ ࠙ࠚࡉࠨࠗ࠻ http://www.nacalai.co.jp ⹜⮎ߪߎߎߦ ଔᩰ⚊ᦼߩߏᾖળ 0 1 2 0 - 4 8 9 - 5 5 2 ຠߦ㑐ߔࠆᛛⴚ⊛ߥߏᾖળ E-mail:[email protected] TEL:075-211-2703 FAX:075-211-2673 0511
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