对于短模板，我们建议增加模板浓度，聚合酶浓度和反应时间，因为小 RNA 的转录在很大程度上取决于起始事件的数量。无 RNase 水：µl 10X 转录缓冲液：4 µl 20X 核糖核苷酸混合溶液：2 µl 模板 (0.3–0.5 µg)：µl 20X HMW混合：2 µl T7 RNA 聚合酶（500 单位/µl）：4 µl 总反应体积：40 µl 在 42°C 下孵育至少 3-4 小时。过夜孵育将导致更高的产量。注意：转录产量在前 90 分钟（大约）内线性增加，并在 2-3 小时后达到最大值。如果需要，反应可以过夜进行，但产量不会更高（转录短于 0.5 kb 的 RNA 时除外）。双链RNA 在 42°C 下长时间孵育后是稳定的。转录反应中使用的模板量取决于纯化方法。对于未纯化的、热灭活的限制性消化物，每 40 µl 反应包含不超过 13 µl 的模板。对于未纯化的 PCR 产物，每 40 µl 反应加入不超过 7 µl。在所有情况下，每 40 µl 反应中添加的模板 DNA 量不应超过 2 µg，因为模板浓度高于此值时 RNA 产量不会更高。虽然 HiScribe T7 In Vitro Transcription Kit 中的所有成分都是 RNase-免费的，可以将核糖核酸酶引入实验室环境的转录反应中。这可以通过以下一些简单的预防措施来避免：1）在处理 RNA 时始终戴上手套，2）使用一套专门用于 RNA 工作的移液器或防气溶胶（屏障）移液器吸头，3）使用超纯水（Milli- Q 或等效物）并高压灭菌所有解决方案，以及 4) 尽可能使用一次性塑料器皿代替玻璃器皿。没有必要用焦碳酸二乙酯 (DEPC) 处理溶液和设备。如果处理后 DEPC 没有完全失活，它可以抑制后续反应。
此资源的链接产品类别：已停产 (HomeProtocols14 分钟转换协议 (C2987H/C2987I)14 分钟转换协议 (C2987H/C2987I)概述与高效转换协议相比，以下协议的效率仅为 25%。对于 C2987H，在提供的试管中执行步骤 1-6。方案对于 C2987H：从 -80°C 冰箱中取出细胞并在手中解冻。对于 C2987I：在冰上解冻一管 NEB 5-alpha 感受态大肠杆菌细胞，直到最后冰晶消失。将 50 µl 细胞轻轻小心混匀，移取置于冰上的转化管中。向细胞混合物中加入 1-5 µl 含有 1 pg-100 ng 质粒 DNA。小心轻弹管子 4-5 次以混合细胞和 DNA。不要涡旋。将混合物置于冰上 10 分钟。不要混合。在 42°C 下热激 30 秒。不要混合。置于冰上 3 分钟。不要混合。将 200 µl 室温 SOC 移入混合物中。立即将 50-100 µl 涂抹到选择板上并过夜孵育战斗温度为 37-42°C。注：使用氨苄西林以外的抗生素进行选择可能需要在选择性培养基上进行电镀之前进行一些生长。菌落在高于 37°C 的温度下发育得更快，但某些构建体在高温下可能不稳定。此资源的链接产品类别：克隆感受态细胞菌株产品应用：USER® 克隆、USER® 和耐热 USER II 酶的应用、转化相关产品：NEB ® 5-alpha 感受态大肠杆菌（高效）家乡和资源搜索通过更新搜索字段，选择过滤器类型并应用其他过滤器来完善您的搜索。
您的搜索返回1328结果。显示：1050100250all«1…12412512612712812812912913131313213333333333搜索结果使用修饰的核苷酸 (E2065)ProtocolmiRNA 检测 (E3312)ProtocolsgRNA Synthesis Using the HiScribe™ Quick T7 High Yield RNA Synthesis Kit (NEB #E2050)ProtocolsiRNA Transfection Protocol for TransPass R1Protocolα-Lytic Protease In-gel Digestion Protocol (NEB #P8113)Protocolα-裂解蛋白酶典型反应方案 (NEB #P8113)方案 α1-3,6 半乳糖苷酶应用说明 (P0731)方案HomeTools & ResourcesSearchSearch通过更新搜索字段、选择过滤器类型和应用其他过滤器来优化您的搜索。
您的搜索返回了 1328 个结果。显示：1050100250All«1…567891011121314…133» 搜索结果TitleTypeCarrier Protein 27 (Array) Protocol 2：通过考马斯蓝染色的 SDS-PAGEProtocolCarrier 蛋白 27（Western）协议 1 进行的结扎分析：对于 Western BlottingProtocol Carrier Protein 27（Western）协议 2：通过考马斯蓝染色的 SDS-PAGEProtocolCarrier 蛋白 39（阵列）协议 I 进行的结扎分析：连接和 ArrayProtocolCarrier Protein 39 (Array) Protocol II：通过考马斯蓝染色的 SDS-PAGEProtocolCarrier Protein 39 (Western) Protocol 1：对于 Western BlottingProtocolCarrier Protein 39 (Western) Protocol 2 的连接分析：通过考马斯蓝染色的 SDS-PAGEProtocolCell 的连接分析LysisProtocolCellular Labeling (E9100)Protocol 细胞标记 (E9120)ProtocolHomeTools & ResourcesSearchSearch通过更新搜索字段、选择过滤器类型和应用其他过滤器来优化您的搜索。
您的搜索返回了 1328 个结果。显示：1050100250All«1…45678910111213…133» 搜索结果TitleTypeCLuc 活性检测方案 II（注射器-配备发光计）（E3314）ProtocolCP 反应缓冲液方案 1：Western BlottingProtocol CP 反应缓冲液方案 2：通过考马斯亮蓝 SDS-PAGE 进行的连接分析 ProtocolCappable-seq 用于原核转录起始位点 (TSS) determinationProtocolCapped RNA Synthesis (E2040)ProtocolCapped RNA Synthesis ( E2050)ProtocolCapping Protocol (M2080)ProtocolCapture 甲基化 CpG DNA (E2600)ProtocolCapture and Elute Step for Control DNA (E2600)ProtocolCarrier Protein 27 (Array) Protocol 1: Ligation and ArrayProtocolHomeTools & ResourcesSearchSearch通过更新搜索字段、选择过滤器类型和应用其他过滤器来优化您的搜索。
您的搜索返回了 1328 个结果。显示：1050100250All«1…3456789101112…133» 搜索结果TitleTypeAppendix for use with the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB #E7760, #E7765) and NEBNext Ultra II RNA Library Prep Kit for Illumina (NEB #E7770, #E7775)ProtocolAssess library quality on a Bioanalyzer®（安捷伦高灵敏度芯片）(E7420)方案在 Bioanalyzer®（安捷伦高灵敏度芯片）(E7530) 上评估文库质量使用 PolyA Spin™ mRNA 分离试剂盒ProtocolBio-P1：将肽连接到硫酯标签蛋白的 C 端ProtocolBlotting and HybNEBlot Phototope Kit 生成的探针的去除。NEB PCR 克隆试剂盒的钝化协议协议CLuc 活性测定协议 I（无注射器的发光计）（E3314）协议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.Hydrophilic Streptavidin Magnetic Beads are 2-3 m superparamagnetic particles covalently coupled to a highly pure form of streptavidin. The beads can be used to capture biotin labeled substrates including antigens, antibodies and nucleic acids.The strength of the biotin-streptavidin interaction, an association constant (Ka) of 1015 M-1 coupled with the low non-specific binding of streptavidin permits captured substrates to be useful as ligands in subsequent experiments including mRNA isolation and the capture of primary or secondary antibodiesBinding capacity: 30 g biotinylated antibody per mg and 400 pmol of single-stranded 25 bp biotinylated oligonucleotide per mgQuick, small-scale purification of biotinylated oligonucleotides affords higher throughput and efficiencyHydrophilic Streptavidin Magnetic Beads are 2 m superparamagnetic particles covalently coupled to a highly pure form of streptavidin. The beads can be used to capture biotin labeled substrates including antigens, antibodies and nucleic acids (1,2). The strength of the biotin-streptavidin interaction, an association constant (Ka) of 1015 M-1 coupled with the low non-specific binding of streptavidin permits captured substrates to be useful as ligands in subsequent experiments including mRNA isolation and the capture of primary or secondary antibodies (3,4).Beads are supplied as a 4 mg/ml suspension in a phosphate buffer (PBS) (pH 7.4) containing 0.1% BSA, 0.02% NaN3 and 0.05% Tween -20.For larger volume requirements, customized and bulk packaging is available by purchasing through the OEM/Bulks department at NEB. Please contactcustom@neb.comfor further information.Reagents SuppliedThe following reagents are supplied with this product:Binding CapacityThe beads will bind greater than 800 pmol of free biotin per mg and greater then 400 pmol of single-stranded 25 bp biotinylated oligonucleotide per mg. Beads will bind greater than 800 pmol of free biotin per mg and greater than 400 pmol of single-stranded 20 bp biotinylated oligonucleotide per mg of Hydrophilic Streptavidin Magnetic Beads. Chung, S. et al. (2005). J. Biol. Chem.. 280, 4578. Tang, B. et al. (2003). Genome. 46, 833--840. Zhang, X. et al. (2006). J. Clin. Invest.. 116, 3050-3059. Fernandez-Cabezudo, M.J. et al. (2004). Internat. Immunol.. 16, 1215-1223. Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.SpecificationsThe Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. 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