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

Citations & References (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
Price: 870.00 EUR

TOPO™ TA Cloning™ Dual Promoter kits are for fast, efficient cloning and subsequent in vitro transcription. These kits include theRead more

Change view

Catalog Number	Quantity
K460001	25 Reactions
K4600J10	10 Reactions
K460040	50 Reactions

3 Options

Catalog number K460001

Price (EUR)

870,00

Each

Add to cart

Quantity:

25 Reactions

Price (EUR)

870,00

Each

Add to cart

TOPO™ TA Cloning™ Dual Promoter kits are for fast, efficient cloning and subsequent in vitro transcription. These kits include the pCR™II-TOPO™ TA vector with dual T7 and SP6 promoters. By eliminating time-consuming and tedious restriction site cloning, TOPO™ cloning is the most reliable cloning method, featuring a 3-step protocol and 5-minute cloning reaction, and yielding up to 95% recombinants.

Convenient features of this TOPO™ TA Cloning™ Dual Promoter kit include:
• 3´-T overhangs for direct ligation of Taq-amplified PCR products
• T7 and SP6 promoters for efficient in vitro transcription
• M13 forward and reverse primer sites for sequencing
• 16 convenient restriction sites, including EcoRI, flanking your insert for subsequent excision or subcloning
• Kanamycin and ampicillin resistance for your choice of selection in E. coli
• Easy blue/white colony screening for selection of recombinants
• Includes OneShot™ TOP10 Competent cells for added convenience and highest cloning efficiencies

Kit Options: TOPO™ TA Cloning™ Dual Promoter Kits
TOPO™ TA Cloning™ Dual Promoter kits can be purchased with a variety of competent cells that deliver different advantages depending upon your needs:
• General cloning: TOP10 cells (Cat. Nos. K4600-J10, K4600-01, K4600-40)
• High-efficiency cloning: TOP10 Electrocomp™ cells (Cat. Nos. K4660-01, K4660-40)
• General cloning, bacteriophage T1 resistance: DH5α-T1R cells (Cat. Nos. K4620-01, K4620-40)
• Fast growth: Mach1™ -T1R chemically competent E. coli (Cat. No. K4610-20)
• Repressor/induction needs: TOP10F’ cells (Cat. Nos. K4650-01, K4650-40)
• Provide your own cells (Cat. Nos. 451641, 450641)

For Research Use Only. Not for use in diagnostic procedures.

Specifications

Bacterial or Yeast StrainTOP10

Cell TypeChemically Competent

Cloning MethodTOPO™-TA

Product LineOne Shot

Product TypeCloning Kit

PromoterT7, SP6

Quantity25 Reactions

VectorTOPO-TA Cloning Vectors

FormatKit

Unit SizeEach

Contents & Storage

Box 1:
• Topoisomerase I-activated pCR™II-TOPO™ vector
• PCR buffer
• Salt solution
• dNTPs
• Control template
• M13 forward and reverse primers
• Control PCR primers
• Sterile water

Store at -5 to -30°C.
All reagents are stable for 6 months when properly stored.

Box 2:
• One Shot™ Chemically Competent or Electrocomp™ E. coli
• S.O.C. medium
• Supercoiled pUC19 control plasmid.

Store at -68 to -85°C.

Frequently asked questions (FAQs)

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