TA Cloning™ 试剂盒,双启动子,含 pCR™II 载体和 One Shot™ INVαF' 化学感受态大肠杆菌
TA Cloning&trade; 试剂盒,双启动子,含 pCR&trade;II 载体和 One Shot&trade; INV&alpha;F' 化学感受态<i>大肠杆菌</i>
Invitrogen™

TA Cloning™ 试剂盒,双启动子,含 pCR™II 载体和 One Shot™ INVαF' 化学感受态大肠杆菌

TA Cloning™ 试剂盒双启动子(含 pCR™II 载体)提供了一种快速的一步法克隆策略,可将 Taq 扩增 PCR了解更多信息
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货号反应次数
K205040
又称 K2050-40
40 次反应
K20500120 次反应
货号 K205040
又称 K2050-40
价格(CNY)
7,026.00
飞享价
Ends: 31-Dec-2025
14,049.00
共减 7,023.00 (50%)
Each
添加至购物车
反应次数:
40 次反应
价格(CNY)
7,026.00
飞享价
Ends: 31-Dec-2025
14,049.00
共减 7,023.00 (50%)
Each
添加至购物车
TA Cloning™ 试剂盒双启动子(含 pCR™II 载体)提供了一种快速的一步法克隆策略,可将 Taq 扩增 PCR 产物直接插入质粒载体中。pCR™II 载体的 T7 和 Sp6 启动子可在体外转录插入片段,产生正义链或反义链产物。TA Cloning 试剂盒双启动子使用 pCR™II 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™ 试剂盒双启动子的特点:
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™II 载体提供:
• 3'-T 突出端,用于直接连接 Taq 扩增 PCR 产物
• T7 和 Sp6 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:含 One Shot™ INVF' 化学感受态大肠杆菌(K2050-01 和 K2050-40)、含 One Shot™ TOP10F' 化学感受态大肠杆菌(K2060-01 和 K2060-40)以及不含感受态细胞(K2750-20 和 K2750-40),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INVαF ́
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
包括线性化 pCRII 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、对照品、One Shot INVαF' 化学感受态大肠杆菌、S.O.C. 培养基和超螺旋对照质粒。
反应次数40 次反应
产品线One Shot
产品类型克隆试剂盒
促进剂T7, SP6
数量40 rxns
载体pCRII
产品规格试剂盒
Unit SizeEach
内容与储存
本试剂盒含有线性化 pCR™II 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)

Have you compared in vitro transcription levels between SP6 and T7 promoters in your pCRII vectors?

No, we have not done in-house comparisons of transcription levels. It is widely known though that T7 polymerase produces more RNA than SP6 (on the order of 10-fold higher).

引用和文献 (9)

引用和文献
Abstract
Semaphorin III can function as a selective chemorepellent to pattern sensory projections in the spinal cord.
Authors:Messersmith EK, Leonardo ED, Shatz CJ, Tessier-Lavigne M, Goodman CS, Kolodkin AL
Journal:Neuron
PubMed ID:7748562
'Distinct classes of primary sensory neurons in dorsal root ganglia subserve different sensory modalities, terminate in different dorsoventral locations in the spinal cord, and display different neurotrophin response profiles. Large diameter muscle afferents that terminate in the ventral spinal cord are NT-3 responsive, whereas small diameter afferents subserving pain and ... More
Kainate receptor subunits expressed in single cultured hippocampal neurons: molecular and functional variants by RNA editing.
Authors:Ruano D, Lambolez B, Rossier J, Paternain AV, Lerma J
Journal:Neuron
PubMed ID:7748549
To determine the kainate receptor subunits that are found in native kainate receptors, we have applied a multiplex PCR of cDNAs reverse transcribed from mRNA harvested from single cultured hippocampal neurons after electrophysiological recording. We found that all the cells showing rapidly desensitizing currents in response to kainate express the ... More
Map-based cloning of chloronerva, a gene involved in iron uptake of higher plants encoding nicotianamine synthase.
Authors:Ling HQ, Koch G, Baumlein H, Ganal MW
Journal:Proc Natl Acad Sci U S A
PubMed ID:10359845
The uptake of iron in plants is a highly regulated process that is induced on iron starvation. In tomato, the mutant chloronerva exhibits constitutive expression of iron uptake responses and intercostal chlorosis. Biochemically, chloronerva is an auxotroph for nicotianamine, a key polyamine in plant iron uptake metabolism. The chloronerva gene ... More
ARIA, a protein that stimulates acetylcholine receptor synthesis, is a member of the neu ligand family.
Authors:Falls DL, Rosen KM, Corfas G, Lane WS, Fischbach GD
Journal:Cell
PubMed ID:8453670
Motor neurons stimulate their postsynaptic muscle targets to synthesize neurotransmitter receptors. Polypeptide signaling molecules may mediate this inductive interaction. Here we report the purification of ARIA, a protein that stimulates the synthesis of muscle acetylcholine receptors, and the isolation of ARIA cDNA. Recombinant ARIA increases acetylcholine receptor synthesis greater than ... More
The accessory subunit of Xenopus laevis mitochondrial DNA polymerase gamma increases processivity of the catalytic subunit of human DNA polymerase gamma and is related to class II aminoacyl-tRNA synthetases.
Authors:Carrodeguas JA, Kobayashi R, Lim SE, Copeland WC, Bogenhagen DF
Journal:Mol Cell Biol
PubMed ID:10330144
Peptide sequences obtained from the accessory subunit of Xenopus laevis mitochondrial DNA (mtDNA) polymerase gamma (pol gamma) were used to clone the cDNA encoding this protein. Amino-terminal sequencing of the mitochondrial protein indicated the presence of a 44-amino-acid mitochondrial targeting sequence, leaving a predicted mature protein with 419 amino acids ... More