PCR Enzyme

High Affinity HotStart Taq

High affinity antibody-modified HotStart DNA polymerase

Catalog number / packaging

Cat. no	ID	No. of preps
4992774	ET108-02	500 U,5 U/ μl
4992773	ET108-01	250 U,5 U/ μl

Manual Inquire

Storage

Store at -20℃.

Store at -20℃.
Description

The High Affinity HotStart Taq in this product is a mixed product of TIANGEN Taq DNA Polymerase and its monoclonal antibody, which is suitable for HotStart PCR experiments. Before the PCR reaction is heated at high temperature, the Taq monoclonal antibody will combine with Taq polymerase to inhibit its polymerase activity. The high affinity antibody in the product can ensure the complete shielding of Taq polymerase activity at room temperature, so that the whole reaction system has high specificity. In addition, Taq DNA Polymerase in this product has higher template affinity, which can improve amplification efficiency and specificity. The 3' end of PCR product produced by the kit is A, which can be directly cloned into TA vector.

本製品の中のHigh Affinity HotStart Taqは当社のTaq DNAPolymerase及びそのモノクローナル抗体の混合物であり、HotStart PCR試験に適しています。PCR反応の高温加熱前にTaqモノクローナル抗体はTaq酵素と結合し、そのポリメラーゼ活性を抑制します。本製品中の高親和力抗体は常温条件下でTaq酵素活性を完全に遮断することを確保し、反応システムの全体に高い特異性を持たせます。また、本製品のTaq DNA Polymeraseは高いテンプレート親和性を持ち、増幅の効率と特異性を向上させます。本キットで発生したPCR生成物3’の末端はAであり、直接TAベクターでクローンすることができます。
Features

■ High affinity system: High Affinity HotStart Taq is a specific antibody-modified HotStart DNA polymerase with high antibody affinity; Meanwhile, Taq DNA Polymerase has high template affinity, stable amplification efficiency and high specificity. The enzyme is matched with a carefully optimized buffer system to ensure the high sensitivity of PCR reaction.

■ Strong stability: This product has a high success rate for experiments that cannot be completed by common DNA polymerases such as complex templates, low copy template PCR reactions and multiple PCR reactions.

■ Wide applications: This product is not only suitable for ordinary PCR analysis, but also suitable for quantitative PCR analysis.

■高親和システム:High Affinity HotStart Taqは特異的な抗体修飾のホットスタートDNAポリメラーゼであり、抗体親和性が高い。同時にTaq DNA Polymeraseは高いテンプレート親和力を持ち、安定した増幅効率と高い特異性を持っている。この酵素は最適化されたバッファーシステムと結び付け、同時にPCR反応に高感度のメリットを持たせる。

■優れた安定性:複雑なテンプレート、低コピー数テンプレートのPCR反応、多重PCR反応などの一般的なDNAポリメラーゼによってできない試験に対しては、本製品は高い反応成功率を持っている。

■幅広い応用:本製品は普通のPCR分析だけでなく、定量PCR分析にも適している。
Applications

Regular PCR, multiplex PCR, dye-based and probe-based real-time PCR.

Regular PCR, multiplex PCR, dye-based and probe-based real-time PCR.
Precautions

1.The 5×Probe qPCR Buffer is only needed for probe-based qPCR, but not for regular PCR dye-based qPCR reaction.

2. When performing probe-based qPCR, good amplification results can be obtained using 250 nM final concentration of primer and 200 nM final concentration of probe in most systems. If it is necessary to further optimize the primer concentration, it is suggested to adjust in the range of 50-900 nM; If the probe concentration needs to be further optimized, adjust in the range of 100-500 nM.

1.The 5×Probe qPCR Buffer is only needed for probe-based qPCR, but not for regular PCR dye-based qPCR reaction.

2. When performing probe-based qPCR, good amplification results can be obtained using 250 nM final concentration of primer and 200 nM final concentration of probe in most systems. If it is necessary to further optimize the primer concentration, it is suggested to adjust in the range of 50-900 nM; If the probe concentration needs to be further optimized, adjust in the range of 100-500 nM.

Experimental Example

Set up Real-Time PCR reaction system by dye method:

Set up Real-Time PCR reaction system by dye method:
Reaction program

* The dosage of ROX Reference Dye depends on different real-time PCR instruments.

Reaction program

* The dosage of ROX Reference Dye depends on different real-time PCR instruments.
In this experiment, SYBR Green fluorescence quantitative reagent configured by High Affinity HotStart Taq is used to perform real-time qPCR detecting GAPDH of total RNA of human Hela cells. cDNA synthesis is carried out using FastQuant RT Kit, and cDNA input is equivalent to 20 ng-2 pg of total RNA.

In this experiment, SYBR Green fluorescence quantitative reagent configured by High Affinity HotStart Taq is used to perform real-time qPCR detecting GAPDH of total RNA of human Hela cells. cDNA synthesis is carried out using FastQuant RT Kit, and cDNA input is equivalent to 20 ng-2 pg of total RNA.
In this experiment, SYBR Green fluorescence quantitative reagent configured by High Affinity HotStart Taq is used to perform real-time qPCR detecting GAPDH of total RNA of human Hela cells. cDNA synthesis is carried out using FastQuant RT Kit, and cDNA input is equivalent to 20 ng-2 pg of total RNA.

In this experiment, SYBR Green fluorescence quantitative reagent configured by High Affinity HotStart Taq is used to perform real-time qPCR detecting GAPDH of total RNA of human Hela cells. cDNA synthesis is carried out using FastQuant RT Kit, and cDNA input is equivalent to 20 ng-2 pg of total RNA.

PCR Enzyme

High Affinity HotStart Taq

Catalog number / packaging

Storage

Description

Features

Applications

Precautions

Experimental Example

Publication