TA Cloning ™试剂盒,含 PCR™2.1 载体,不含感受态细胞
TA Cloning ™试剂盒,含 PCR™2.1 载体,不含感受态细胞
Invitrogen™

TA Cloning ™试剂盒,含 PCR™2.1 载体,不含感受态细胞

TA 克隆™试剂盒(含 PCR™2.1 载体)提供了一种快速的一步法克隆策略,可将经 Taq 扩增的 PCR 产物直接插入质粒载体中了解更多信息
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货号反应次数
K20202020 次反应
K20204040 次反应
货号 K202020
价格(CNY)
2,278.00
飞享价
Ends: 31-Dec-2025
4,553.00
共减 2,275.00 (50%)
Each
添加至购物车
反应次数:
20 次反应
价格(CNY)
2,278.00
飞享价
Ends: 31-Dec-2025
4,553.00
共减 2,275.00 (50%)
Each
添加至购物车
TA 克隆™试剂盒(含 PCR™2.1 载体)提供了一种快速的一步法克隆策略,可将经 Taq 扩增的 PCR 产物直接插入质粒载体中。TA Cloning™ 试剂盒使用 pCR™2.1 克隆载体和 ExpressLink™ T4 DNA 连接酶,在十五分钟的室温连接步骤中生成连接产物。反应通常产生 >80% 包含插入片段的重组体。

TA Cloning™ 试剂盒(含 pCR™2.1 载体)的特点:
快速且方便 — 15分钟、室温连接
高效 — 蓝/白斑筛选和 >80% 含正确插入片段的克隆体
灵活 — 可选择卡那霉素或氨苄青霉素抗性,实现灵活的抗生素选择
轻松 — 无需对 PCR 产物进行任何酶修饰
流程简化 — 不需要使用包含限制性位点的 PCR 引物

pCR™2.1 载体提供:
• 3'-T 突出端,用于直接连接 Taq 扩增 PCR 产物
• T7 启动子,用于体外 RNA 转录和测序
• 两侧具有 EcoR I 位点的通用性多位点接头,可方便地切除插入片段
• M13 正向和反向引物位点,用于测序

TA Cloning™ 的工作原理
Taq 聚合酶具有非模板依赖性活性,可在 PCR 产物的 3' 末端引入单脱氧腺苷 (A)。该试剂盒中提供的线性化载体具有单个 3' 脱氧胸苷 (T) 残基。这使得 PCR 插入片段高效地与载体连接。

试剂盒配置
TA Cloning™ 试剂盒提供多种配置:不含感受态细胞(K2020-20 和 K2020-40)、含 One Shot™ INVF' 化学感受态大肠杆菌(K2000-01 和 K2000-40)、含 One Shot™ TOP10F' 化学感受态大肠杆菌(K2030-01 和 K2030-40)以及含 One Shot™ TOP10 化学感受态大肠杆菌(K2040-01 和 K2040-40),分 20 和 40 次反应试剂盒规格。
仅供科研使用。不可用于诊断程序。
规格
细菌或酵母菌株不包括在内
克隆方法TA克隆
适用于(应用)PCR克隆
反应次数20 次反应
产品线TA Cloning™
产品类型克隆试剂盒
促进剂T7
数量20 reactions
载体pCR2.1
产品规格试剂盒
Unit SizeEach
内容与储存
TA Cloning™ 试剂盒包含线性化 pCR™2.1 载体、ExpressLink™ T4 DNA 连接酶、5X ExpressLink™ T4 DNA 连接缓冲液、dNTP、10X PCR 缓冲液、无菌水和对照品。

将所有组成部分储存在 -20°C 下。妥善储存时,所有试剂均可保证 6 个月的稳定。

常见问题解答 (FAQ)

How do you recommend that I prepare my DNA for successful electroporation of E. coli?

For best results, DNA used in electroporation must have a very low ionic strength and a high resistance. A high-salt DNA sample may be purified by either ethanol precipitation or dialysis.

The following suggested protocols are for ligation reactions of 20ul. The volumes may be adjusted to suit the amount being prepared.

Purifying DNA by Precipitation: Add 5 to 10 ug of tRNA to a 20ul ligation reaction. Adjust the solution to 2.5 M in ammonium acetate using a 7.5 M ammonium acetate stock solution. Mix well. Add two volumes of 100 % ethanol. Centrifuge at 12,000 x g for 15 min at 4C. Remove the supernatant with a micropipet. Wash the pellet with 60ul of 70% ethanol. Centrifuge at 12,000 x g for 15 min at room temperature. Remove the supernatant with a micropipet. Air dry the pellet. Resuspend the DNA in 0.5X TE buffer [5 mM Tris-HCl, 0.5 mM EDTA (pH 7.5)] to a concentration of 10 ng/ul of DNA. Use 1 ul per transformation of 20 ul of cell suspension.

Purifying DNA by Microdialysis: Float a Millipore filter, type VS 0.025 um, on a pool of 0.5X TE buffer (or 10% glycerol) in a small plastic container. Place 20ul of the DNA solution as a drop on top of the filter. Incubate at room temperature for several hours. Withdraw the DNA drop from the filter and place it in a polypropylene microcentrifuge tube. Use 1ul of this DNA for each electrotransformation reaction.

When should DMSO, formamide, glycerol and other cosolvents be used in PCR?

Cosolvents may be used when there is a failure of amplification, either because the template contains stable hairpin-loops or the region of amplification is GC-rich. Keep in mind that all of these cosolvents have the effect of lowering enzyme activity, which will decrease amplification yield. For more information see P Landre et al (1995). The use of co-solvents to enhance amplification by the polymerase chain reaction. In: PCR Strategies, edited by MA Innis, DH Gelfand, JJ Sninsky. Academic Press, San Diego, CA, pp. 3-16.

Additionally, when amplifying very long PCR fragments (greater than 5 kb) the use of cosolvents is often recommended to help compensate for the increased melting temperature of these fragments.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.