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TOPcloner™ TA core Kit

No Product Name Cat.# Conc Size Price(₩) Add
1 TOPcloner™ TA core Kit EZ011S - 20 reactions ₩ 200,000 추가
2 TOPcloner™ TA core Kit EZ011M - 40 reactions ₩ 360,000 추가
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Product description

TOPcloner™ TA core kit is suitable for cloning Taq polymerase-amplified PCR products with A-tail overhangs at room temperature in 5 min. The vector provided in this kit contains topoisomerase covalently coupled to its ends, which allows efficient cloning of DNA fragments with A-tail overhangs to the vector without using a regular ligase.

 

Characteristics

- Short reaction time (5 min)

- Highly efficient cloning

- Direct use of PCR products

 

Applications

- Fast and efficient cloning of A-tailed PCR products (for example, Taq-amplified DNA)

 

Supplied with

- pTOP TA V2

- 6X TOPcloner™ Buffer

- Sterile water

 

Quality control

- Cloning efficiency: 99%

 

Standard reaction conditions

6X TOPcloner™ Buffer

1 μl

pTOP TA V2 (10 ng/μl)

1 μl

PCR product

1~4 μl

Sterile water

up to 6 μl

Incubate at room temperature(25) for 5 min

Transformation




Figure 1. A schematic map of pTOP TA V2 (3807 bp).
A molecule of topoisomerase (TOPO) is covalently coupled to each of the two 3' one-nt protruding ends (nt positions 294 and 295) of pTOP TA V2 (Cat.# EZ001 and EZ011). Therefore, pTOP TA V2 is ready to form a covalent bond when mixed with PCR products with one-nt 3' A overhang. The addition of one A to the duplex DNA ends is intrinsic to Taq DNA polymerase.

 - Lac promoter/operator : 95-216
 - M13 Reverse Primer binding site : 205-221
 - LacZ ORF : 217-534
 - MCS, Multiple Cloning Sites : 234-357
 - M13 (-20) Forward Primer binding site : 391-406
 - ccdB ORF : 544-846
 - Kanr gene : 1057-1989
 - Ampr gene : 2007-2867
 - pUC origin : 3012-3685

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