SafeView取代EtBr |新型核酸安全染劑ABM G108
新型 安全核酸染劑
SafeView™/Safedye是一種可代替溴化乙錠(EtBr)的新核酸染料,其為新一代安全性螢光染劑,靈敏度與EtBr相當。
核酸觀察箱 (UV/Blue)須知
一般核酸觀察箱分為UV與藍光二種系統;傳統的UV Box觀察箱, 具有長波(365nm)、中波(312nm)、短波(256nm)波段激發。長波能量較弱 (365nm) ,適合切膠等膠體回收(gel extraction)用。短波能量強(256nm),可以取得較明顯的影像。新一型Safe Blue藍光觀察箱,取代UV的波長為470nm 藍光波段,減少UV光線對樣品及眼睛造成的傷害。搭配不同的核酸觀察箱,abmgood提供不同波長之核酸染劑,將核酸影像最佳化。也減少做實驗誘癌之風險發生。
傳統的核酸染劑, EtBr (Biovisioin #1203-10)
溴化乙錠 (縮寫為EtBr),是一種核酸染劑,用為瓊脂糖凝膠電泳後,核酸之染色。在UV紫外光照射下,未結合的ErBr可被激發出橙紅色螢光;結合的DNA或雙股RNA,螢光強度會增強20倍,使得核酸電泳後可分辨核酸相對的分子量。EtBr其激發(Excitation)波段為310nm,散射光(emmision)為570nm。需注意溴化乙錠 (EtBr)為是一種強的誘變劑intercalator,可致癌或致畸。操作時必須佩戴手套。
G108-G 操作手冊 MSDS Ligation效能測試報告
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新型安全核酸染劑, SafeView/Safedye
SafeView/Safedye安全染劑,產品經由Ames test測試,証明對生物體無致癌性。產品激發(Excitation)波段為290~490nm,散射光(Emmision)為480~605nm。不同染劑呈現不同激發(Excitation)及散射光(Emmision),依使用方式分為二種型式Safeview(內或外染)及safedye(直接當loading dye使用)。請選擇合的波長範圍,才能將影像最佳化
SafeViewTM系列產品:Classic/FireRed/Plus (內染或外染)
SafeView™ -EtBr替代品 (內染或外染使用)。使用方法與EtBr類似,無須更改使用習慣,可使用藍光/UV偵測。
SafeView™ Classic-dsDNA與ssDNA可呈現綠色螢光、RNA呈現紅色螢光。 SafeView™ FireRed-dsDNA、ssDNA、以及RNA呈現紅色螢光。 SafeView Plus™-作為外染使用。不需退染步驟。靈敏度更佳。
SafedyeTM系列產品:Safe-Green、Red、White、Pack (loading dye染劑)
Safe dyes直接跑電泳時, 當loading dye使用。是一種快速染核之loading dye,不需將膠體預染或外染。
靈敏度比較 (核酸安全染劑 V.S. EtBr)
傳統EtBr染劑,其靈敏度是檢測至少1ng以下的DNA;新型安全染劑,具有更高的signal:Bkgrd的比值,靈敏度高於傳統型EtBr染劑。在選擇不同染劑前,也請選擇適的波長範圍,才能將影像最佳化。
Safe-Green has Excitation Wavelength of 490nm and Emission Wavelength of 525nm, and its sensitivity range is between 0.2-0.6ng.
Safe-Red has Excitation Wavelength of 540nm and Emission Wavelength of 630nm, and its sensitivity range is between 0.3-0.8ng.
Safe-White has Excitation Wavelength of 370nm and Emission Wavelength of 470nm, and its sensitivity range is between 0.2-0.5ng.
SafeView Classic emits green fluorescence when bound to dsDNA and ssDNA and red fluorescence when bound to RNA. This stain has one excitation (490 nm) and two emission spectra (520 nm and 635 nm) and the sensitivity range is between 0.1-0.3ng.
SafeView Plus has Excitation Wavelength of 490nm and Emission Wavelength of 525nm and its sensitivity range is between 0.05-0.1ng.
核酸安全染劑產品應用:
可使用在瓊脂糖agarose或丙烯醯胺凝膠polyacrylamide gels中的 dsDNA、ssDNA及RNA。搭配不同波長濾片進行DNA偵測。In gel stining、Gel extraction、Ligation、Transformation、Transfection
核酸安全染劑使用方法:
SafeView 產品100ml agarose溶液中加入5ul SafeView™ Classic 後製膠, Safedye為跑電泳時將maker與樣本中加入SafeView™ dye (1:5倍稀釋)
(內染) SafeView™ Classic、FireRed:100ml agarose溶液中加入5μl SafeView™ 後進行製膠。
(外染) SafeView Plus™:DNA電泳完成後,將膠體泡入含有SafeView Plus™的電泳緩衝液(1:10,000稀釋),室溫下反應10-40分鐘即可進行偵測。
(是loading dye也是DNA染劑) SafeView™ dyes:將maker/樣本中加入SafeView™ dye(1:5稀釋)後進行DNA電泳即可。
G108 does have 2 emission wavelengths, but only one excitation. G108-G has only one excitation and one emission wavelength.
Please refer to the following information to help clarify the emission and excitation spectrum of all our SafeView products:
Safe-Green (G108-G) has Excitation Wavelength of 490nm and Emission Wavelength of 525nm, and its sensitivity range is between 0.2-0.6ng.
Safe-Red (G108-R) has Excitation Wavelength of 540nm and Emission Wavelength of 630nm, and its sensitivity range is between 0.3-0.8ng.
Safe-White (G108-W) has Excitation Wavelength of 370nm and Emission Wavelength of 470nm, and its sensitivity range is between 0.2-0.5ng.
SafeView (G108) Classic emits green fluorescence when bound to dsDNA and ssDNA and red fluorescence when bound to RNA. This stain has one excitation (490 nm) and two emission spectra (520 nm and 635 nm) and the sensitivity range is between 0.1-0.3ng.
SafeView Plus (G468) has Excitation Wavelength of 490nm and Emission Wavelength of 525nm and its sensitivity range is between 0.05-0.1ng.
產品編號 | 產品名稱 | 波長(Ex/Em) | 靈敏度(ng) | 包裝 |
SafeView™ Classic-內染 | ||||
G108 | SafeView™ Classic |
490nm/ 635nm 490nm/520nm |
0.1-0.3 | 1ml |
SafeView Plus™-外染 | ||||
G468 | SafeView Plus™ | 290nm/520nm | 0.05-0.1 | 1ml |
SafeView™ dyes | ||||
G108-G | Safe-Green™ | 490nm/525nm | 0.2-0.6 | 1ml |
G108-R | Safe-Red™ | 490nm/630nm | 0.3-0.8 | 1ml |
G108-W | Safe-White™ | 370nm/470nm | 0.2-0.5 | 1ml |
G108-P | Safe-Pack™ | 參照G108-G、R、W |
安全測試報告 (Ligation Efficiency Test Report)
核酸染劑測試報告,為比較安全染劑Safe-GreenTM與溴化乙錠( Ethidium Bromide)使用相比,Safe-GreenTM對連接反應( ligation reactions)是否有負面影響
To investigate if Safe-GreenTM has any negative effect on ligation reactions compared to using Ethidium Bromide.
效能測試操作程序
1)使用XhoI線性化(Linearize ) 6 µg載體(vector)。
2)將3 µg線性化載體 (linearized vector)/ DNA Loading dye /溴化乙錠(Ethidum Bromide )凝膠上運行; 另外在沒有溴化乙錠取3 µg線性化的載體+ SafeGreen (含Loading dye) )凝膠上運行。
3)使用Column-Pure DNA凝膠提取試劑盒,以25 µl洗脫緩衝液(elution buffer)洗脫DNA。
4)使用4 µl純化的線性化載體和T4 DNA連接酶建立連接反應, 設置no ligase控件以評估未切割的載體背景的水平,如下所示:
5 ) 轉化(Transform)至LB +氨芐青黴素平板(ampicillin plates)上以選擇含有所需的重組質粒。第二天比較菌落計數。
安全染劑(Safe-Green)與傳統溴化乙錠相比,使用Safe-Green時,連接效率沒有差異
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