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TOPcloner™ Blunt core Kit

No Product Name Cat.# Conc Size Price(₩) Add
1 TOPcloner™ Blunt core Kit EZ012S - 20 reactions ₩ 200,000 추가
2 TOPcloner™ Blunt core Kit EZ012M - 40 reactions ₩ 360,000 추가
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Product description

TOPcloner™ Blunt core kit is suitable for cloning Pfu polymerase-amplified PCR products with blunt ends at room temperature in 5 min. The vector provided in this kit contains topoisomerase covalently coupled to its ends, which allows efficient cloning of blunt-ended DNA fragments 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 blunt-ended PCR products (for example, Pfu-amplified DNA)

 

Supplied with

- pTOP Blunt V2

- 6X TOPcloner™ Buffer

- Sterile water

 

Quality control

 - Cloning efficiency: 99%

 

Standard reaction conditions

6X TOPcloner™ Buffer

1 μl

pTOP Blunt 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. A schematic map of pTOP Blunt V2 (3807 bp).
A molecule of topoisomerase (TOPO) is covalently coupled to the 3' blunt duplex ends (nt positions 294 and 295) of pTOP Blunt V2 (Cat.# EZ002 and EZ012). Therefore, pTOP Blunt V2 can form a covalent bond with blunt-ended PCR products obtained by Pfu 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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