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

Zitierungen und Referenzen (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
Price: 1.373.65 EUR

Ansicht ändern

Katalognummer	Menge
K460040	50 Reaktionen
K4600J10	10 Reaktionen
K460001	25 Reaktionen

3 Optionen

Katalognummer K460040

Preis (EUR)

1.373,65

Exklusiv online

1.416,00

Ersparnis 42,35 (3%)

Each

Zum Warenkorb hinzufügen

Menge:

50 Reaktionen

Preis (EUR)

1.373,65

Exklusiv online

1.416,00

Ersparnis 42,35 (3%)

Each

Zum Warenkorb hinzufügen

Die TOPO™ TA Cloning™ Dual Promoter Kits ermöglichen schnelles, effizientes Klonen und anschließende In-vitro-Transkription. Diese Kits enthalten den pCR™II-TOPO™ TA-Vektor mit dualen T7- und SP6-Promotern. Durch die Eliminierung des zeitaufwändigen und langwierigen Klonens von Restriktionsstellen ist die TOPO™-Klonierung die zuverlässigste Methode, die mit einem 3-Schritt-Protokoll nur 5 Minuten in Anspruch nimmt und bis zu 95 % Rekombinanten liefert.

Zu den praktischen Funktionen dieses TOPO™ TA Cloning™ Dual Promoter Kits gehören:
• 3´-T-Überhänge für die direkte Ligation Taq-amplifizierter PCR-Produkte
• T7- und SP6-Promoter für effiziente In-vitro-Transkription
• M13 Vorwärts- und Rückwärtsprimer-Stellen für die Sequenzierung
• 16 praktische Restriktionsstellen, einschließlich EcoRI, zu beiden Seiten Ihres Inserts für nachfolgende Exzision oder Subklonierung
• Kanamycin- und Ampicillin-Resistenz für Ihre individuelle Auswahl an E. coli
• Leichtes Blau-Weiß-Kolonie-Screening zur Auswahl von Rekombinanten
• Enthält OneShot™ TOP10 Kompetente Zellen für zusätzlichen Komfort und höchste Klonierungseffizienz

Kit-Optionen: TOPO™ TA Cloning™ Dual Promoter Kits
TOPO TA Cloning Dual Promoter Kits sind mit einer Vielfalt kompetenter Zellen erhältlich, die verschiedene Vorteile entsprechend Ihren Anforderungen bieten:
• Allgemeine Klonierung: Top10-Zellen (Kat.-Nr. K4600-J10, K4600-01, K4600-40)
• Hocheffiziente Klonierung: TOP10 Electrocomp™ Zellen (Kat.-Nr. K4660-01, K4660-40)
• Klonierung allgemein, Resistenz gegen Bakteriophagen T1: DH5α-T1R-Zellen (Kat.-Nr. K4620-01, K4620-40)
• Schnelles Wachstum: Mach1™-T1R chemisch kompetente E. coli (Bestell-Nr. K4610-20)
• Repressor/Induktion benötigt: Top10F’-Zellen (Bestell-Nr. Nr. K4650-01, K4650-40)
• Ihre eigenen Zellen (Bestell-Nr. 451641, 450641)

Nur für Forschungszwecke. Nicht zur Verwendung bei diagnostischen Verfahren.

Specifications

Bakterien- oder HefenstammTOP10

ZelltypChemically Competent

KlonierungsmethodeTopo™-TA

ProduktlinieOne Shot

ProdukttypKlonierungskit

PromoterT7, SP6

Menge50 Reaktionen

VektorTOPO TA-Klonierungsvektoren

FormatKit

Unit SizeEach

Inhalt und Lagerung

Box 1:
• Topoisomerase-I-aktivierter pCR™II-TOPO™ Vektor
• PCR-Puffer
• Salzlösung
• dNTPs
• Kontroll-Template
• M13-Vorwärts- und Rückwärtsprimer
• Kontroll-PCR-Primer
• steriles Wasser

Bei -5 bis -30 °C lagern.
Alle Reagenzien sind bei ordnungsgemäßer Lagerung 6 Monate lang stabil.

Box 2:
• One Shot™ chemisch kompetente oder Electrocomp™ E. coli
• S.O.C.-Medium
• Superhelikales pUC19-Kontrollplasmid

Lagerung bei -68 bis -85 °C.

Häufig gestellte Fragen (FAQ)

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

Zitierungen und Referenzen (8)

Zitierungen und Referenzen

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