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BioDynamics Laboratory Inc.(株式会社バイオダイナミクス研究所)
BioDynamics Laboratory Inc.
[Date:October 28 2014]

More efficient and fast cloning of PCR products DynaExpress Hetero-Stagger PCR Cloning Kit

TA cloning of PCR product is known as much less efficient cloning method than not only cohesive end but also blunt end cloning method.
DynaExpress Hetero-Stagger PCR Cloning Kit enables highly efficient and fast cloning of PCR products.
This novel method does not require any enzymatic procedures such as restriction enzyme, ligase, exonuclease, Urasil DNA glycosylase and Cre-loxP recombinase reactions.

Instead, the method needs four PCR primers. Two PCR primers contain eight or nine extra bases at the 5' ends with target sequence depending on non-proofreading or proofreading thermostable DNA polymerase.
The other two primers have only target sequence. Two PCR reactions are set up to generate two PCR products containing different extra terminal sequences.

The two PCR products and a compatible vector, pHST, are mixed, heat-denatured and annealed to form a heteroduplex product with nine bases overhangs complementary to the single-strand extensions of pHST vector.
As the complementary extension of the vector and insert are long, and ligation reaction is not required.
The mixture of annealed vector and PCR products can be used directly for the transformation of chemical competent cells.

Features

  • High ligation efficiency.
  • Simple principle.
  • Simple procedure (mix, heat and anneal)
  • Does not require any enzymatic reaction
  • Direct use for transformation after annealing
  • Choice of insert orientation
  • Applicable to both proofreading and non-proofreading DNA polymerase
  • Total time from PCR product to plating is just 90 mins.

Principle

DynaExpress Hetero-Stagger PCR Cloning Kit Principle

Comparison Data

DynaExpress Hetero-Stagger PCR Cloning Kit Principle

The effect on cloning using different amounts of PCR fragments (about 1kb) was tested regarding the DynaExpress Hetero-Stagger PCR Cloning Kit and Supplier A's TA cloning Kit. The recommended protocol from each supplier was followed. After the reaction between PCR fragments and vector, competent cells were transformed with 1/3 amounts of reaction mixture.
Half of the amounts of the cells were plated onto the agar plate. The white bars and the purple bars show the numbers of white colonies.
The blue bars show the numbers of blue colonies.

Supplier Product Code Product Name Size Price
BDLバイオダイナミクス
BioDynamics Laboratory Inc.(株式会社バイオダイナミクス研究所)

Datasheet

DS158

Hetero-Stagger PCR Cloning Kit, DynaExpress

1kit

$265
Supplier Product Code Product Name Size Price
BDLバイオダイナミクス
BioDynamics Laboratory Inc.(株式会社バイオダイナミクス研究所)

DS158

Hetero-Stagger PCR Cloning Kit, DynaExpress

1kit

$265
Description

Hetero-Stagger PCR Cloning Kit enables highly efficient and fast cloning of PCR products. This novel method does not require any enzymatic procedures. pHST Vector is included.

Storage-20°CCAS
LinkDynaExpress Hetero-Stagger PCR Cloning Kit

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