TA Cloning™试剂盒,含 pCR™2.1 载体和 One Shot™ TOP10F' 化学感受态大肠杆菌
TA Cloning&trade;试剂盒,含 pCR&trade;2.1 载体和 One Shot&trade; TOP10F' 化学感受态<i>大肠杆菌</i>
Invitrogen™

TA Cloning™试剂盒,含 pCR™2.1 载体和 One Shot™ TOP10F' 化学感受态大肠杆菌

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
K20300120 次反应
K20304040 次反应
货号 K203001
价格(CNY)
3,809.00
飞享价
Ends: 31-Dec-2025
7,618.00
共减 3,809.00 (50%)
Each
添加至购物车
反应次数:
20 次反应
价格(CNY)
3,809.00
飞享价
Ends: 31-Dec-2025
7,618.00
共减 3,809.00 (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TOP10F ́
细胞类型化学感受态大肠杆菌
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One Shot
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。感受态细胞试剂盒包含 One Shot™ 化学感受态大肠杆菌、S.O.C. 培养基和一种超螺旋对照质粒。

在 -80°C 下储存 One Shot™ 大肠杆菌。所有其他组分储存在 -20°C 下。正确存放时,所有试剂均可保证 6 个月的稳定。

常见问题解答 (FAQ)

我可以使用TOP10F’感受态细胞对包含ccdB基因的TOPO载体进行转化吗?

含F质粒的菌株,如TOP10F’,不推荐用于转化和筛选任何含ccdB基因的TOPO载体的重组克隆。F质粒编码CcdA蛋白,该蛋白是CcdB旋转酶-毒素蛋白的抑制剂,但是CcdA蛋白的半衰期比CcdB蛋白短。当没有足够的CcdA抑制CcdB蛋白时,过表达CcdB蛋白会引起细胞死亡。

Can I use TOP10F' competent cells for transformation of my TOPO vector that contains the ccdB gene?

Strains that contain an F plasmid, such as TOP10F', are not recommended for transformation and selection of recombinant clones in any TOPO vector containing the ccdB gene. While the F plasmid does encode the CcdA protein, which acts as an inhibitor of the CcdB gyrase-toxin protein, the half-life of the CcdA protein is shorter than that of the CcdB protein. Overexpression of the CcdB protein causes cell death when its action is not prevented by sufficient CcdA.

引用和文献 (4)

引用和文献
Abstract
Xath5 participates in a network of bHLH genes in the developing Xenopus retina [published erratum appears in Neuron 1998 Nov;21(5):following 1221]
Authors:Kanekar S, Perron M, Dorsky R, Harris WA, Jan LY, Jan YN, Vetter ML
Journal:Neuron
PubMed ID:9390513
'We examined the function of basic-helix-loop-helix (bHLH) transcription factors during retinal neurogenesis. We identified Xath5, a Xenopus bHLH gene related to Drosophila atonal, which is expressed in the developing Xenopus retina. Targeted expression of Xath5 in retinal progenitor cells biased the differentiation of these cells toward a ganglion cell fate, ... More
T cell receptor recognition of MHC class II-bound peptide flanking residues enhances immunogenicity and results in altered TCR V region usage.
Authors:Carson RT, Vignali KM, Woodland DL, Vignali DA
Journal:Immunity
PubMed ID:9324359
'Naturally processed MHC class II-bound peptides possess ragged NH2 and COOH termini. It is not known whether these peptide flanking residues (PFRs), which lie outside the MHC anchor residues, are recognized by the TCR or influence immunogenicity. Here we analyzed T cell responses to the COOH-terminal PFR of the H-2A(k) ... More
High-performance subtractive hybridization of cDNAs by covalent bonding between specific complementary nucleotides [In Process Citation]
Authors:Ying SY, Lin S
Journal:Biotechniques
PubMed ID:10337490
We have developed an improved subtractive hybridization method that provides a fast, simple and reliable isolation of desired different sequences from two compared DNA libraries, one of which contains all unwanted homologues (subtracter) and another contains certain desired heterologues (tester). The DNA library can be made from either mRNA or ... More
Immunogene therapy of tumors with vaccine based on Xenopus homologous vascular endothelial growth factor as a model antigen.
Authors: Wei Y Q; Huang M J; Yang L; Zhao X; Tian L; Lu Y; Shu J M; Lu C J; Niu T; Kang B; Mao Y Q; Liu F; Wen Y J; Lei S; Luo F; Zhou L Q; Peng F; Jiang Y; Liu J Y; Zhou H; Wang Q R; He Q M; Xiao F; Lou Y Y; Xie X J; Li Q; Wu Y; Ding Z Y; Hu B; Hu M; Zhang W;
Journal:Proc Natl Acad Sci U S A
PubMed ID:11553767
Overcoming immune tolerance of the growth factors associated with tumor growth should be a useful approach to cancer therapy by active immunity. We used vascular endothelial growth factor (VEGF) as a model antigen to explore the feasibility of the immunogene tumor therapy with a vaccine based on a single xenogeneic ... More