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TArget CloneTM -Plus-
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TArget CloneTM/
TArget CloneTM -Plus-Code No. TAK-101 10 reactions / 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.
Details
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
- Mizuguchi H, Nakatsuji M, Fujiwara S, Takagi M and Imanaka T, J Biochem., 126: 762-8 (1999)