TOPO™ TA Cloning™ Kit, Dual Promoter, with One Shot™ TOP10 chemically competent E. coli cells

TOPO™ TA Cloning™ Kit, Dual Promoter, with One Shot™ TOP10 chemically competent <i>E. coli</i> cells

Citas y referencias (8)

Invitrogen™

TOPO™ TA Cloning™ Kit, Dual Promoter, with One Shot™ TOP10 chemically competent E. coli cells

Name: TOPO™ TA Cloning™ Kit, Dual Promoter, with One Shot™ TOP10 chemically competent E. coli cells
SKU: K460040

Los kits de promotores dobles TOPO™ TA Cloning™ se han concebido para la clonación rápida y eficaz y la posteriorMás información

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Número de catálogo	Cantidad
K460040	50 reacciones
K4600J10	10 reacciones
K460001	25 reacciones

3 opciones

Número de catálogo K460040

Precio (MXN)

Cantidad:

50 reacciones

Los kits de promotores dobles TOPO™ TA Cloning™ se han concebido para la clonación rápida y eficaz y la posterior transcripción in vitro. Estos incluyen el vector pCR™II-TOPO™ TA con promotores dobles de T7 y SP6. Al eliminar la tediosa y laboriosa clonación de sitios de restricción, la clonación TOPO™ es el método más fiable, ya que solo tarda 5 minutos utilizando un protocolo de 3 pasos y produce recombinantes de hasta un 95 %.

Entre las prácticas características del kit de promotor doble TOPO™ TA Cloning™ se incluyen:
• Salientes 3´-T para la ligadura directa de productos de PCR amplificados con Taq
• Promotores de T7 y SP6 para una transcripción in vitro eficaz
• Sitios de primers directos e inversos M13 para secuenciación o detección de PCR
• 16 prácticos sitios de restricción, incluido EcoRI, que flanquean su inserto para su posterior escisión o subclonación
• Diversas opciones de resistencia a kanamicina y ampicilina en E. coli
• Fácil detección de colonias azules/blancas para la selección de recombinantes
• Incluye células competentes OneShot™ TOP10 para una mayor comodidad y una máxima eficacia de la clonación

Opciones del kit: Kits de promotor doble TOPO™ TA Cloning™
Los kits de promotor doble TOPO™ TA Cloning™ pueden adquirirse con diversas células competentes que ofrecen distintas ventajas dependiendo de sus necesidades:
• Clonación general: Células TOP10 (n.º de cat. K4600-J10, K4600-01, K4600-40)
• Clonación de alta eficiencia: Células TOP10 Electrocomp™ (n.º de cat. K4660-01, K4660-40)
• Clonación general, resistencia T1 bacteriófaga: Células DH5α-T1R (n.º de cat. K4620-01, K4620-40)
• Crecimiento rápido: E. coli Mach1™ -T1R químicamente competente (n.º de cat. K4610-20)
• Necesidades de represor/inducción: Células TOP10F’ (n.º de cat. K4650-01, K4650-40)
• Proporcione sus propias células (n.º de cat. 451641 y 450641).

Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.

Especificaciones

Cepa bacteriana o de levaduraTOP10

Tipo de célulaQuímicamente competente

Método de clonaciónTOPO™-TA

Línea de productosOne Shot

Tipo de productoKit de clonación

PromotorT7, SP6

Cantidad50 reacciones

VectorVectores TOPO-TA Cloning

FormatoKit

Unit SizeEach

Contenido y almacenamiento

Caja 1:
• Vector pCR™II-TOPO™ activado con la topoisomerasa I
• Tampón de PCR
• Solución salina
• dNTP
• Plantilla de control
• Primers directos e inversos M13
• Primers de PCR de control
• Agua estéril

Almacenar entre -5 y -30 °C.
Todos los reactivos son estables durante 6 meses si se almacenan adecuadamente.

Caja 2:
• E. coli químicamente competente One Shot™ o Electrocomp™
• Medio S.O.C.
• Plásmido de control superhelicoidal pUC19

Almacenar a entre -68 y -85 °C.

Preguntas frecuentes

Can I store my competent E. coli in liquid nitrogen?

We do not recommend storing competent E. coli strains in liquid nitrogen as the extreme temperature can be harmful to the cells. Also, the plastic storage vials are not intended to withstand the extreme temperature and may crack or break.

How should I store my competent E. coli?

We recommend storing our competent E. coli strains at -80°C. Storage at warmer temperatures, even for a brief period of time, will significantly decrease transformation efficiency.

What is the difference between the pCR2.1-TOPO and pCR4-TOPO vectors?

The vector backbones for both of these vectors are very similar. The main difference is that the pCR4-TOPO vector has sequencing primer sites located as close as 33 base pairs from the PCR product insertion site. This minimizes the amount of vector DNA sequence that needs to be read before reaching the sequence of the insert, making the pCR4-TOPO vector very useful for sequencing applications.

What is the difference between the pCR2.1-TOPO and pCRII-TOPO vectors?

The vector backbones for both of these vectors are very similar. The main difference is that the pCRII-TOPO vector is a dual promoter vector, containing the SP6 and T7 promoters for in vitro transcription/sequencing, whereas the pCR2.1-TOPO vector contains only the T7 promoter for in vitro transcription/sequencing. Both vectors contain the M13 Forward and Reverse primer sites for sequencing or PCR screening.

Your TOPO cloning kits contain a control template and control primers. Can I obtain the sequence of the control template?

The sequence of the control template is proprietary.

View all TOPO™ TA Cloning™ Kit, Dual Promoter, with One Shot™ TOP10 chemically competent E. coli cells, 50 Reactions FAQs

Citations & References (8)

Citations & References

Abstract

Cloning and functional expression of a thyrotropin receptor cDNA from rat fat cells.

Authors:Endo T; Ohta K; Haraguchi K; Onaya T;

Journal:J Biol Chem

PubMed ID:7738021

Thyrotropin receptor (TSH-R) has been thought to be thyroid-specific, but, by Northern blot analysis, we found that rat adipose tissue expressed TSH-R mRNAs in amounts approaching those in the thyroid. To investigate the function of TSH-R from adipose tissue, we screened a rat fat cell lambda gt11 cDNA library for ... More

Dual DNA binding specificity of ADD1/SREBP1 controlled by a single amino acid in the basic helix-loop-helix domain.

Authors:Kim JB, Spotts GD, Halvorsen YD, Shih HM, Ellenberger T, Towle HC, Spiegelman BM

Journal:Mol Cell Biol

PubMed ID:7739539

'Adipocyte determination- and differentiation-dependent factor 1 (ADD1), a member of the basic helix-loop-helix (bHLH) family of transcription factors, has been associated with both adipocyte differentiation and cholesterol homeostasis (in which case it has been termed SREBP1). Using PCR-amplified binding analysis, we demonstrate that ADD1/SREBP1 has dual DNA sequence specificity, binding ... More

Sequential mutations in the interleukin-3 (IL3)/granulocyte-macrophage colony-stimulating factor/IL5 receptor beta-subunit genes are necessary for the complete conversion to growth autonomy mediated by a truncated beta C subunit.

Authors:Hannemann J, Hara T, Kawai M, Miyajima A, Ostertag W, Stocking C

Journal:Mol Cell Biol

PubMed ID:7739524

'An amino-terminally truncated beta C receptor (beta C-R) subunit of the interleukin-3 (IL3)/granulocyte-macrophage colony-stimulating factor/IL5 receptor complex mediates factor-independent and tumorigenic growth in two spontaneous mutants of a promyelocytic cell line. The constitutive activation of the JAK2 protein kinase in these mutants confirms that signaling occurs through the truncated receptor ... More

A powerful transgenic tool for fate mapping and functional analysis of newly generated neurons.

Authors:Zhang J, Giesert F, Kloos K, Vogt Weisenhorn DM, Aigner L, Wurst W, Couillard-Despres S

Journal:BMC Neurosci

PubMed ID:21194452

Lack of appropriate tools and techniques to study fate and functional integration of newly generated neurons has so far hindered understanding of neurogenesis' relevance under physiological and pathological conditions. Current analyses are either dependent on mitotic labeling, for example BrdU-incorporation or retroviral infection, or on the detection of transient immature ... More

Classical and non-classical Major Histocompatibility Complex class I gene expression in in vitro derived bovine embryos.

Authors:Doyle J, Ellis SA, O'Gorman GM, Aparicio Donoso IM, Lonergan P, Fair T

Journal:J Reprod Immunol

PubMed ID:19682752

The role of the Major Histocompatibility Complex class I (MHC-I) genes in human and mouse preimplantation embryo development has received considerable attention. In contrast, information concerning the role of these genes in bovine embryo development is limited. The objective of the current study was to characterize the expression pattern of ... More

View all TOPO™ TA Cloning™ Kit, Dual Promoter, with One Shot™ TOP10 chemically competent E. coli cells, 50 Reactions citations