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yT&A Cloning Kit

 

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yT&A Cloning Kit

The yT&A cloning kit contain the yT&A vector and many reagents needed for ligation. It is a convenient pack for cloning PCR product generated using thermostable DNA polymerases, such as YEAtaq polymerase, which add a single terminal 3’-dA nucleotide overhang. After ligation, the mixture can be used directly to transform into competent cells (ECOSTM perform better) or be purified first to achieve higher transformation efficiency. The kit contains enough reagents for 20 reactions.

 

 


 

Features and Benefits

Fast!! Ligation can be completed within 5 ~ 20 minutes vs. 1 hour to overnight by using similar systems from other companies.
Higher transformation efficiency in comparison to other competitor’s cloning system.
More accurate results in comparison to other companies in colony pickings.
Two types of ligation buffer provided for your convenience.


 

Applications

Accepts terminal 3’-dA nucleotide overhang PCR products
Transform ligation product (purified/unpurified) into ECOSTM or competent cells
LacZ complementation for blue/white screening
Ampicillin marker for antibiotic selection


 

Storage Conditions

Store at –20°C


 

yT&A Cloning Vector(download cloning vector sequence)

With ampicillin resistance
With blue/white screening capability
All pass rigorous QC test to ensure the background colonies are close to zero
Very stable

Figure: Map and Sequence reference points of the yT&A cloning vector

Before the insert incorporated into the yT&A cloning vector, there is only 1 HindIII site and no BglII site. After the incorporation, the T and A nucleotides on the insert complement the sequence on the vector and generate these two new sites. This merit of yT&A vector makes cloning more economic and convenient.

Position of important sequences:

Multiple Cloning region

434 to 490

lac Z gene

511 to 149

Ampr gene

2528 to 1671

T7 promoter

402 to 439

M13 forward primer

359 to 375

M13 reverse primer

528 to 507

 

Restriction enzyme sites of yT&A cloning vector


 

Experimental Data

The result of ligation using the control insert DNA and reagents provided in yT&A cloning kit.

* The size of colony PCR on the agarose gel will change according to the primer design. In this colony PCR, the size of the band on the gel is

 

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Last modified: 05/29/09