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TArget Clone/ TArget Clone -Plus-

<TArget Clone> Code No. TAK-101 10 reactions

<TArget Clone -Plus-> Code No. TAK-201 20 reactions


Description

TArget Clone and TArget Clone -Plus- are high efficient TA cloning kits. TArget Clone [Code No. TAK-101] can be applied to the TA cloning of PCR products amplified using Taq DNA polymerase, Blend Taq [Code No. BTQ-101], Blend Taq -Plus- [Code No. BTQ-201] or KOD Dash [Code No. LDP-101]. Almost all PCR products having dA overhanging at the 3'end are available. TArget Clone -Plus- [Code No. TAK-201] can be applied to the TA cloning of blunt-end PCR products amplified using KOD -Plus- [Code No. KOD-201] or KOD FX [Code No. KFX-101]. TArget Clone -Plus- contains 10xA-attachment mix. This reagent is a mixture of anti-KOD DNA polymerase antibody specific to KOD 3'→5' exonuclease activity (proof-reading activity) and Taq DNA polymerase, which exhibits terminal transferase activity. The 10 x A-attachment mix allows for blunt end PCR products to acquire overhanging dA at the 3'-ends.


Product Name Applicable for PCR products amplified with
TArget Clone Taq DNA polymerase, Blend Taq, Blend Taq -Plus-, KOD Dash <PCR products having overhanging dA at the 3'-ends>
TArget Clone -Plus- KOD -Plus-, KOD FX


Fig.1 Principle of 10x A-attachment mix


Fig.2 Map of pTA2 vector


Features

    [TArget Clone & TArget Clone -Plus-]
  • - Ligation step can be completed in 5 min.
  • - PCR products can be used without purification.
  • - The vector generates blue E. coli colonies on X-gal/(IPTG) plates in case of no inserts.

    [TArget Clone -Plus-]
  • - Realizes TA cloning of blunt-end products from KOD -Plus- and KOD FX.
  • - Addition of dA to the 3'-ends of PCR product can be competed in 10 min at 60ºC.

Applications

    [TArget Clone]
  • TA cloning of PCR products amplified using Taq DNA polymerase, Blend Taq [Code No. BTQ-101], Blend Taq -Plus- [Code No. BTQ-201] or KOD Dash [Code No. LDP-101]. Almost all PCR products having dA overhanging at the 3'end are available.

    [TArget Clone -Plus-]
  • TArget Clone -Plus- [Code No. TAK-201] can be applied to the TA cloning of blunt-end PCR products amplified using KOD -Plus- [Code No. KOD-201] or KOD FX [Code No. KFX-101].

Storage condition

Store at -20ºC


Components

[TArget Clone]

pTA2 Vector(50ng/µl) 10 µl
2x Ligation Buffer 50 µl
T4 DNA Ligase 10 µl

[TArget Clone -Plus-]

pTA2 Vector(50ng/µl) 10 µl
2x Ligation Buffer 50 µl
T4 DNA Ligase 10 µl
10x A-attachment Mix 10 µl

Application data

Example 1. TA cloning of a 500 bp-PCR product amplified with KOD- Plus- and Blend Taq.

Human b-globin gene (0.5 kb) was amplified using KOD -Plus- [Code No. KOD-201] or Blend Taq [BTQ-101]. Then, the unpurified PCR products were cloned into the pTA2 vectors using TArget Clone or TArget Clone -Plus- according to each instruction manual. Subsequently, DH5a competent cells were transformed using the ligation products and cultured on the LB/Amp/X-gal plate overnight at 37ºC. As shown Fig. 1, PCR products from Blend Taq were successfully cloned into the pTA2 Vector using both kits. And, PCR products from KOD -Plus- was cloned efficiently into pTA2 Vector using TArget Clone -Plus-.

Fig. 1 Comparison of cloning efficiency

References

1) Mizuguchi H, Nakatsuji M, Fujiwara S, Takagi M and Imanaka T, J Biochem., 126: 762-8 (1999)