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TOPcloner™ XL TA Kit new

No Product Name Cat.# Conc Size Price(₩) Add
1 TOPcloner™ XL TA Kit EZ033S - 10 reactions ₩ 210,000 추가
2 TOPcloner™ XL TA Kit EZ033M - 20 reactions ₩ 378,000 추가
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Product description

TOPcloner™ XL TA Kit is suitable for cloning Taq polymerase-amplified long (3-10 kb) 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 ligation of DNA fragments with A-tail overhangs to the vector without using a regular ligase. This kit provides special gel purification reagents that ensures efficient cloning of long, full-length PCR products.

 

Characteristics

- Highly efficient long PCR product (3-10 kb) cloning

- Short reaction time (5 min)

- Direct use of PCR products

 

Applications

- Fast and efficient cloning of long, full-length (3-10 kb) A-tailed PCR products.

 

Component

Box 1

pTOP TA V2, 6X TOPcloner buffer, M13 reverse primer, M13 forward primer Control insert,

Control primers mix, Sterile water

Box 2

S.O.C. Media, DH5α chemical competent E.coli, pUC19 control DNA

Box 3

Buffer GB, Buffer NW, Buffer EB, Crystal Violet, 6XCrystal Violet Loading Buffer

 

Standard reaction conditions

6X TOPcloner™ buffer

1 µl

pTOP TA V2 (10 ng/µl)

1 µl

PCR products

1-4 µl

Distilled water

up to 6 µl

→ Incubate at room temperature (25) for 5 min

→ Transform the ligated DNA into competent E.coli cells.

 

 


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.


 

 

Figure 2. The result of colony PCR.

Large (10 kb) size of PCR product was cloned using TOPcloner™ XL TA Kit.

● Material Safety Data Sheet

- Enzynomics_MSDS_EZ033_TOPcloner™ XL TA Kit_E


● Vector Information

1. Vector full sequence

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