TOPO™ TA, Dual TOP10 (Supply Center Packaging)

The TOPO™ TA Cloning™ Dual Promoter Kit is for fast, efficient cloning and subsequent in vitro transcription/translation. This kit includesRead more

Catalog Number	Quantity
K460001SC also known as K4600-01SC	25 Reactions

Catalog number K460001SC

also known as K4600-01SC

Price (HKD)

Quantity:

25 Reactions

The TOPO™ TA Cloning™ Dual Promoter Kit is for fast, efficient cloning and subsequent in vitro transcription/translation. This kit includes the pCR™II-TOPO™ TA vector (see figure). By eliminating time-consuming and tedious restriction site cloning, TOPO™ cloning is the most reliable method, taking only 5 minutes using a 3-step protocol, and yielding up to 95% recombinants. Convenient features of the 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

This product comes in Supply Center packaging, which features an additional outer box that holds both the TOPO™ vector box and the competent cells box. This special packaging helps ensure that visitors to a Supply Center obtain the complete kit.

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

Specifications

Bacterial or Yeast StrainTOP10

Cloning MethodTOPO™-TA

Product LineOne Shot

Product TypeCloning Promoter Kit

Quantity25 Reactions

Shipping ConditionDry Ice

VectorTOPO-TA Cloning Vectors

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, Dual TOP10 (Supply Center Packaging) FAQs

Citations & References (1)

Citations & References

Abstract

Multiple sites of V lambda diversification in cattle.

Authors:Lucier MR, Thompson RE, Waire J, Lin AW, Osborne BA, Goldsby RA

Journal:J Immunol

PubMed ID:9820519

'Ig repertoire diversification in cattle was studied in the ileal Peyer''s patch (IPP) follicles of young calvesand in the spleens of late first-trimester bovine fetuses. To investigate follicular diversification, individualIPP follicles were isolated by microdissection; VA diversity was examined by RT-PCR and subsequentcloning and sequencing. When 52 intrafollicular sequences from ... More