Invitrogen™ TOPO™ TA Cloning™ Kit for Sequencing, with pCR™4-TOPO™ Vector, One Shot™ TOP10 Chemically Competent E. coli, and PureLink™ Quick Plasmid Miniprep Kit

Description

Includes

Topoisomerase I-activated pCR™4-TOPO™ vector; PCR buffer; Salt solution; dNTPs; Control template; T3, T7, M13 forward, and M13 reverse primers; Control PCR primers; Sterile water; One Shot™ Chemically Competent or Electrocomp™ E. coli; S.O.C. medium; Supercoiled pUC19 control plasmid

Provides a 5 minute, one-step cloning strategy. Invitrogen™ TOPO™ TA Cloning™ Kit for Sequencing, with pCR4-TOPO Vector, One Shot™ TOP10 Chemically Competent E. coli and PureLink Plasmid Prep Kit enables direct insertion of Taq polymerase-amplified PCR products into a plasmid vector for sequencing.

The TOPO™ TA Cloning™ Kits for Sequencing use the pCR™4-TOPO™ TA vector with specially designed sequencing primer sites that return more insert sequence and less vector sequence from each reaction. These kits include everything necessary to clone and select recombinant vectors containing the PCR fragment of choice.

• Get More Sequence—Allows for more insert sequence and less vector sequence when using standard sequencing primers
• Fast and Easy—Go from PCR-to-clones in just 3 steps and in as little as 5 minutes hands-on time
• Efficient—Achieve up to 95% clones with correct insert
• Proven—Reliable performance for over a decade with over 4000 citations

TOPO™ TA Cloning™ Kits for Sequencing Kit Overview

• VECTOR: pCR™4-TOPO™ TA Vector—Optimized cloning vector for improved sequencing results
• CLONING METHOD: TOPO™ TA Cloning —Topoisomerase I based 5 minute ligation of PCR products with 3´A overhangs (Taq-amplified) to the vector
• COMPETENT CELLS: Various Options—Choose from kits with either general, high-efficiency, bacteriophage T1-resistant, or fast-growing competent cells

pCR™4-TOPO™ TA Vector—Optimized for Sequencing
Much of the multiple cloning site from the pCR™4-TOPO™ TA vector has been removed to shorten the distance between sequencing primer sites and the insert site to as little as 33bp. This means sequencing reactions give less vector sequence and more insert sequence. The pCR™4-TOPO™ TA vector has sites for 4 common sequencing primers: M13 forward, M13 reverse, T7, and T3. The kits include an aliquot of each.

pCR™4-TOPO™ TA Clone Selection and Manipulation
The pCR™4-TOPO™ TA vector contains both ampicillin and kanamycin resistance markers and a LacZα-ccdB gene fusion for positive selection and blue/white screening. The vector’s minimized multiple cloning site still includes flanking EcoRI sites for simplified excision of cloned PCR products and a unique Sse8387I site for generation of nested deletions prior to sequencing. T7 and T3 promoters are also present for in vitro transcription.

Simplified TOPO™-Based Cloning
Using TOPO™ cloning technology, there is no need for PCR primers containing specific sequences, post-PCR procedures, vector preparation, or other time-intensive DNA manipulation steps. Just add your PCR reaction straight to the provided topoisomerase-charged vector, incubate 5 minutes, and transform with the providedE. coli competent cells.

Efficient Cloning
With up to 95% of clones carrying the desired insert, you can screen less clones to help you save time and money. The pCR™4-TOPO™ TA vector used in this kit comes with 3´T overhangs for efficient ligation of Taq-amplified PCR products with 3´A overhangs.

The Standard in Cloning
When it comes to cloning, TOPO™ cloning technology has been a reliable partner for thousands of scientists for over ten years. Fast, simple-to-use, and efficient, TOPO™ cloning has been applied to many different vectors for a wide array of applications. The TOPO™ TA Cloning™ Kit for Sequencing can be purchased with a variety of competent cells that deliver different advantages depending on your needs.

Specifications

• Bacterial or Yeast Strain: TOP10
• Cell Type: Chemically Competent
• Cloning Method: TOPO™-TA
• Content And Storage: Box 1:
  • Topoisomerase I-activated pCR™4-TOPO™ vector
  • PCR buffer
  • Salt solution
  • dNTPs
  • Control template
  • T3, T7, M13 forward, and M13 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.
  
  Box 3:
  PureLink Quick Plasmid Miniprep Kit (Cat. no. K4575-02)
  
  Store at room temperature.
• Format: Kit
• No. of Reactions: 25 Reactions
• Product Line: One Shot™
• Product Type: Cloning Kit
• Promoter: T7, T3
• Quantity: 25 Reactions
• Vector: TOPO-TA Cloning Vectors

Safety and Handling

WARNING: Reproductive Harm - www.P65Warnings.ca.gov

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