High Efficiency Transformation Protocol (C3013) | NEB

来自 : 发布时间：2024-04-28

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您的搜索返回了 30 个结果。显示：10All«123» 搜索结果TitleTypeTroubleshooting Guide for CloningTroubleshooting GuideTroubleshooting Guide for DNA Cleanup and Plasmid纯化故障排除指南 末端修饰故障排除指南 基因组 DNA 提取和纯化故障排除指南 (NEB #T3010)故障排除指南 连接酶故障排除指南 LunaScript RT Master Mix（无引物）试剂盒故障排除指南 (NEB #E3025)故障排除指南故障排除帝王蝶 HMW DNA 提取指南试剂盒（NEB #T3050 和 #T3060）故障排除指南NEBExpress® MBP 融合和纯化系统故障排除指南（NEB #E8201）故障排除指南NEBNext® Ultra™ II FS DNA 文库制备试剂盒故障排除指南故障排除指南NEBNext® 故障排除指南Ultra™ II 和 Ultra DNA 文库制备试剂盒故障排除指南主页工具和资源选择图表钝化选择图表钝化选择图表T4 DNA 聚合酶*(NEB #M0203) DNA 聚合酶 I，大 (Klenow) 片段(NEB #M0210)快速钝化试剂盒(NEB #E1201) 绿豆核酸酶(NEB #M0250)应用去除 3´ 悬垂✓✓ ✓✓去除 5´ 悬垂✓填充 5´ 悬垂✓✓✓* T4 DNA 聚合酶具有很强的 3´→ 5´ 外切活性A-tailing Selection Chart首页工具和资源选择图表A-tailing Selection ChartKlenow Fragment(3\'→5\' exo-)(NEB #M0212) Taq DNA PolymeraseFEATURES反应温度37°C75°C加热灭活75°C, 20分钟NoNucleotide cofactorATPdATPHomeTools & ResourcesSearchSearch

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您的搜索返回了 64 个结果。显示：1050All«1234567» 搜索结果TitleTypeProtein Expression and Purification Selection ChartSelection ChartProtein Kinase Substrate RecognitionSelection ChartPurification微珠、色谱柱和树脂选择图表RNA 帽模拟选择图表选择图表RNA 连接酶选择图表选择图表RNA 聚合酶选择图表选择图表RNA 合成试剂盒选择图表可切割平末端选择图表可切割填充5\' 悬垂选择图表按应用推荐的HiScribe RNA 合成试剂盒选择图表HomeTools & ResourcesSearchSearch

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您的搜索返回了 64 个结果。显示：1050All«1234567» 搜索结果TitleTypeIsoschizomersSelection ChartNEB Diluent and Buffer TableSelection ChartNEBNext® Multiplex Oligos Selection图表选择图表OneTaq® 缓冲液推荐选择图表PUREExpress® 与 NEBExpress® 应用图表选择图表磷酸酶选择图表选择图表DNA 修复酶和结构特异性内切核酸酶的特性选择图表DNA 和 RNA 连接酶的特性选择图表外切核酸酶和非特异性内切酶的特性选择图表蛋白酶选择图表选择图表Accept cookiesWe use cookies to understand how you use our site and to improve the overall user experience. This includes personalizing content and advertising. To learn more and manage cookies, please refer to our Cookie Statement.Are you doing COVID-19 related research? Our latest RUO kit, the Luna SARS-CoV-2 RT-qPCR Multiplex Assay Kit, enables high throughput workflows for real-time detection of SARS-CoV-2 nucleic acid using hydrolysis probes. For simple, visual assay results, the SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit includes a color-changing pH indicator for detection of SARS-CoV-2 nucleic acid amplificationLearn about our tools that are helping researchers develop diagnostics and vaccines for the SARS-CoV-2 virus.Product Price ListDownload a PDF containing pricing for our full product list.Free ShippingSave time and money by placing an order with NEB. Take advantage of free shipping for any order totaling over $350. Place your order before 7:30pm EST for overnight delivery.For shipping questions click here.ProtocolsHigh Efficiency Transformation Protocol (C3013)For C3013H: Thaw a tube of T7 Express lysY/Iq Competent E. coli cells on ice for 10minutes. For C3013I: Thaw a tube of T7 Express lysY/Iq Competent E. coli cells on ice until the last ice crystals disappear. Mix gently and carefully pipette 50 l of cells into a transformation tube on ice.Add 1-5 l containing 1 pg-100 ng of plasmid DNA to the cell mixture. Carefully flick the tube 4-5 times to mix cells and DNA.Donotvortex. Place the mixture on ice for 30 minutes. Do not mix.Heat shock at exactly 42 C for exactly 10 seconds. Do not mix.Place on ice for 5 minutes. Do not mix.Pipette 950 l of room temperature SOC into the mixture.Place at 37 C for 60 minutes. Shake vigorously (250 rpm) or rotate.Warm selection plates to 37 C.Mix the cells thoroughly by flicking the tube and inverting, then perform several 10-fold serial dilutions in SOC.Spread 50-100 l of each dilution onto a selection plate and incubate overnight at 37 C. Alternatively, incubate at 30 C for 24-36 hours or at 25 C for 48 hours.Transformation Protocol Variables:Thawing:Cells are best thawed on ice and DNA added as soon as the last bit of ice in the tube disappears. Cells can also be thawed by hand, but warming above 0 C will decrease the transformation efficiency.Incubation of DNA with Cells on Ice:For maximum transformation efficiency, cells and DNA should be incubated together on ice for 30 minutes. Expect a 2-fold loss in transformation efficiency for every 10 minutes this step is shortened.Heat Shock:Both the temperature and the timing of the heat shock step are important and specific to the transformation volume and vessel. Using the transformation tube provided, 10 seconds at 42 C is optimal.Outgrowth:Outgrowth at 37 C for 1 hour is best for cell recovery and for expression of antibiotic resistance. Expect a 2-fold loss in transformation efficiency for every 15 minutes this step is shortened. SOC gives 2-fold higher transformation efficiency than LB medium; and incubation with shaking or rotating the tube gives 2-fold higher transformation efficiency than incubation without shaking.Plating:Selection plates can be used warm or cold, wet or dry without significantly affecting the transformation efficiency. However, warm, dry plates are easier to spread and allow for the most rapid colony formation.* Ideally, DNA for transformation should be purified and resuspended in water or TE. However, up to 10 l of DNA directly from a ligation mix can be used with only a two-fold loss of transformation efficiency. Where it is necessary to maximize the number of transformants (e.g. a library), a purification step, either a spin column or phenol/chloroform extraction and ethanol precipitation should be added.If you don t see your country above, please visit ourinternational siteSession ExpiredYou have been idle for more than 20 minutes, for your security you have been logged out. Please sign back in to continue your session.Sign InYour profile has been mapped to an Institution, please sign back for your profile updates to be completed.Sign InTo save your cart and view previous orders, sign in to your NEB account. Adding products to your cart without being signed in will result in a loss of your cart when you do sign in or leave the site.