产品说明
Product overview Technical/Spec Support/Documents FAQs Tips for usage Application references Related products Description The G-HiFi™ DNA Polymerase is a new genetically modified, recombinant DNA polymerase suitable for GC-rich templates that are difficult to amplify. The fidelity of G-HiFi™ DNA Polymerase is 70 times higher than that of Taq DNA polymerase. The high extension rate of G-HiFi™ DNA Polymerase is achieved by blending the DNA polymerase with an elongation enhancer. The optimized 5X G-HiFi™ Buffer includes special ingredients that suppress non-specific amplification as well as plateau effect produced by conventional PCR. With the optimized 5X G-HiFi™ Buffer, G-HiFi™ DNA Polymerase is capable to amplify most templates, such as longer targets (up to 40 kb from lambda DNA) and that contain GC-rich sequences. Features5’→3’ DNA polymerase activity 3’→5’ exonuclease (proofreading) activity High reaction rate: 7 seconds/kb High fidelity: 70 times higher than Taq polymerase Generates blunt end amplicons Vast elongation capability (up to 40 kb) Thermo-stable for more than 10 hrs at 95°C Storage[TF3000] G-HiFi™ DNA Polymerase-20°C for 24 monthsElongation capabilityG-HiFi™ DNA Polymerase’s high processability enables reliable amplification of λDNA up to 40 kb in length (M: DM5100). SensitivityG-HiFi™ DNA Polymerase performs higher sensitivity for high GC content templates (GC: 71%) compare to high fidelity DNA Polymerase from Brand A (M: DM2000). [TF3000] G-HiFi™ DNA PolymeraseContentsComponentVolumeG-HiFi™ DNA Polymerase (1 U/μl)100 μl5X G-HiFi™ Buffer1200 μldNTPs Mix (2 mM each)600 μlStorage Buffer50 mM Tris-HCl (pH 8.0), 50 mM KCl, 0.1 mM EDTA, 1 mM DTT, stabilizer, 50% (v/v) glycerolUnit DefinitionOne unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid-insoluble material in 30 minutes at 74°C. Storage-20°C for 24 months [TF3000] G-HiFi™ DNA PolymeraseManualManual_TF3000_G-HiFi™ DNA PolymeraseSDSSDS_TF3000 What are the differences between SMO-HiFi™ (TF1000) and G-HiFi™ (TF3000)? SMO-HiFi™ (TF1000) and G-HiFi™ (TF3000) contain different high fidelity DNA polymerases. SMO-HiFi™ (TF1000) and G-HiFi™(TF3000) DNA polymerases are suitable for GC-rich templates that are difficult to amplify. Moreover G-HiFi™ (TF3000) is blended with elongation enhancer to perform quick and accurate PCR. Recommended PCR Condition[TF3000] G-HiFi™ DNA PolymeraseTemplate1 – 150 ngForward primer0.1 – 0.5 µM*Reverse primer0.1 – 0.5 µM*5X G-HiFi™ Buffer10 µldNTPs (2 mM each)5 µlG-HiFi™ DNA Polymerase0.5 – 1 unit**H2Oto 50 µlTotal volume50 µl*When amplifying products ≧ 10 kb in length, use primers at a final concentration of 0.1 μM each.** When amplifying products ≦ 2 kb in length, use 0.5 unit of G-HiFi™ DNA Polymerase. Recommended PCR ProgramFor ≦ 10 kb productsSelect primers with a Tm value of ≧55°C. 20- to 25-mer primers are suitable, or those greater than 25-mer in length may provide optimal results.StepsTemp.TimeCyclesTemplate denature98°C2 min1Denature98°C10 sec25-40Annealing50-68°C15 secExtension68°C10-30 sec/kbFinal extension68°C1 min1 *Optimal PCR condition varies according to primers’ thermodynamic properties.For ≧ 10 kb productsSelect primers with a Tm value of ≧ 65°C. 25- to 35-mer primers are suitable. Avoid high GC-content at the 3" end of each primer.StepsTemp.TimeCyclesDenature98°C10 sec25-40Extension68°C10-30 sec/kb For GC-rich templates: StepsTemp.TimeCycles Templatedenature98°C2 min1 Denature98°C10 sec25-40 Extension68°C10-30 sec/kb AFEAP cloning: a precise and efficient method for large DNA sequence assemblyFanli Zeng,1 Jinping Zang,1 Suhua Zhang,2 Zhimin Hao,1 Jingao Dong,corresponding author1 and Yibin Lincorresponding author3BMC Biotechnol. 2017; 17: 81. Published online 2017 Nov 14. doi: 10.1186/s12896-017-0394-xPMCID: PMC5686892Gel electrophoresisStaining amplicons with safe fluorescent dyes, following by observation under blue-light illuminator to minimize damage of DNA amplicons and maximize successful cloning efficiency. Safe fluorescent dyes[NS1000] FluoroVue™ Nucleic Acid Gel Stain (10,000X), 500 μl[DS1000] FluoroStain™ DNA Fluorescent Staining Dye (Green, 10,000X), 500 μl[DL5000] FluoroDye™ DNA Fluorescent Loading Dye (Green, 6X), 1 mlBlue-light illuminator[VE0100] B-BOX™ Blue Light LED Epi-illuminatorLigationBlund-end PCR amplicons can directly ligate with PCR cloning vector. [CV1100] GetClone™ PCR Cloning Vector II, 20 RxnTransformationPrepare competent cells with high efficiency and transform with time-saving protocol.[CK1000] Champion™ E. coli Transformation KitColony PCRAnalyze colonies with PCR master mix to save preparation time. [TP1100] ExcelTaq™ 5X PCR Master Mix, 200 Rxn[TP1200] ExcelTaq™ 5X PCR Master Dye Mix, 200 Rxn
SMOBIO的B-BOX™是一种长波长蓝光LED落射照明器。它设计紧凑,结构坚固。B-BOX™落射照明器由于其非紫外线光源和低至12伏的低工作电压,并且为非致癌DNA /蛋白质染料提供了工作能力,因此为用户提供了前所未有的安全性。B-BOX™带有高质量的非闪烁LED灯,对DNA和蛋白质染料高度敏感。B-BOX™可以极大地减轻凝胶提取的日常琐事,即使在明亮的环境光下也可以轻松进行可视化和凝胶切割。多角度滤板(目录号VE0102)为凝胶切割,可视化和记录提供了最佳角度。B-BOX™还带有用于过滤和可视化的琥珀色滤镜(目录号VE0103)。Phox™Photobox(目录号 VE0104)为使用任何智能手机拍摄的图像提供了一个迷你暗室环境。最后,工作区域周围的内置屏障系统有助于清洁。
特征
提高克隆效率
紧凑,轻巧和便携式(重量不足1千克)
安全功能包括:470 nm长波长,对用户没有任何紫外线辐射危害
与非致癌,非溴化乙锭的DNA染料兼容
用户友好:样品易于观察(使用滤板或护目镜时)
LED光源持续长达50,000小时
出色的检测灵敏度:使用FluoroStain™DNA荧光染色染料时,DNA≤0.04 ng,使用FluoroStain™蛋白荧光染色染料时,蛋白≤3ng(与银染一样灵敏)
可调和可移动的滤板可进行凝胶切割,可视化和记录
内置屏障设计,易于清理
在明亮的环境光下可见
借助其自身的内置散热器,强调了最小的功率依赖,低发热
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