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 (49)

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: K460001

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
K460001	25 reacciones
K4600J10	10 reacciones
K460040	50 reacciones

3 opciones

Número de catálogo K460001

Precio (MXN)

Cantidad:

25 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

Cantidad25 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, 25 Reactions FAQs

Citations & References (49)

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

Modulation of mouse Paneth cell alpha-defensin secretion by mIKCa1, a Ca2+-activated, intermediate conductance potassium channel.

Authors: Ayabe Tokiyoshi; Wulff Heike; Darmoul Dalila; Cahalan Michael D; Chandy K George; Ouellette Andre J;

Journal:J Biol Chem

PubMed ID:11724775

'Paneth cells in small intestinal crypts secrete microbicidal alpha-defensins in response to bacteria and bacterial antigens (Ayabe, T., Satchell, D. P., Wilson, C. L., Parks, W. C., Selsted, M. E., and Ouellette, A. J. (2000) Nat. Immunol. 1, 113- 38). We now report that the Ca(2+)-activated K(+) channel mIKCa1 modulates ... More

Variant forms of alpha-fetoprotein transcripts expressed in human hematopoietic progenitors. Implications for their developmental potential towards endoderm.

Authors: Kubota Hiroshi; Storms Robert W; Reid Lola M;

Journal:J Biol Chem

PubMed ID:12006569

'Hematopoietic stem cells have been identified as multipotent cells that give rise to all adult hematopoietic lineages. Although the hematopoietic lineage is derived from the mesodermal germ layer in the embryo, recent data suggest that bone marrow cells with an antigenic profile consistent with that of hematopoietic stem cells can ... More

Mutation of the matrix metalloproteinase At2-MMP inhibits growth and causes late flowering and early senescence in Arabidopsis.

Authors: Golldack Dortje; Popova Olga V; Dietz Karl-Josef;

Journal:J Biol Chem

PubMed ID:11726650

'This study characterizes the expression and functional significance of the member of the matrix metalloproteinase (MMP) family At2-MMP from Arabidopsis. By transcript analysis, expression of At2-MMP was found in leaves and roots of juvenile Arabidopsis and leaves, roots, and inflorescences of mature flowering plants showing strong increase of transcript abundance ... More

Phosphorylation of the Carboxyl Terminus of Inner Centromere Protein (INCENP) by the Aurora B Kinase Stimulates Aurora B Kinase Activity.

Authors: Bishop John D; Schumacher Jill M;

Journal:J Biol Chem

PubMed ID:12048181

'How the events of mitosis are coordinated is not well understood. Intriguing mitotic regulators include the chromosomal passenger proteins. Loss of either of the passengers inner centromere protein (INCENP) or the Aurora B kinase results in chromosome segregation defects and failures in cytokinesis. Furthermore, INCENP and Aurora B have identical ... More

View all TOPO™ TA Cloning™ Kit, Dual Promoter, with One Shot™ TOP10 chemically competent E. coli cells, 25 Reactions citations