Sanshibio Taq Hot-Start DNA Polymerase

Taq Hot-Start DNA Polymerase

編號：EH001 規格：500U/1000U/5000U存儲: 儲存於-20°C

產品介紹：

This product is an antibody-blocked hot-start enzyme that suppresses non-specific amplification caused by primer misannealing or primer dimer formation at low temperatures. It is suitable for Hot Start 聚合酶鍊式反應. Since the antibody is inactivated during the initial DNA denaturation step of the PCR reaction, there is no need for special deactivation treatment; it can be used under standard PCR conditions. The PCR product amplified using this product has an ‘A’ base added to the 3′ 結尾, making it directly clonable into a T-Vector. It can also be cloned into blunt-end vectors after end polishing and phosphorylation.

產品內容：

貯存:

儲存於-20°C, 保存期限至少為 12 月.

活動定義:

Using activated mahi-mahi sperm DNA as template/primer, the activity is defined as 1 單元 (U) of acid-insoluble material incorporated, by taking up 10 nmol of nucleotides within 30 minutes at 74°C.

品質管制:

This product has undergone quality testing and is free from deoxyribonuclease endonuclease activity, deoxyribonuclease exonuclease activity, 和核糖核酸酶污染. Host genomic DNA residual content is below 10 副本.

Product Uses:

Hot Start PCR; DNA序列測定.

使用說明:

• Allow the required reagents to equilibrate at room temperature until fully dissolved. 輕輕攪拌均勻 (do not vortex), and use after brief centrifugation to prevent excessive bubble formation and avoid repeated freeze-thaw cycles. If frequently used, store at 4°C. Prepare the PCR reaction mixture according to the following components (prepare the reaction mixture on an ice box):

推薦反應系統:

筆記: 反應體系中各組分的用量可依實際需求調整.

• In general, a two-step method can be used for the reaction; if the two-step amplification is not satisfactory, a three-step method can be used to set up the PCR reaction program.

筆記: 反應條件可依實際要求進行調整和最佳化.

• After the reaction is complete, analyze the experimental results. For detailed analysis methods, refer to the PCR amplification instrument operating manual.

預防措施:

1. Please choose an appropriate annealing (擴大) temperature based on the primer design. 通常, the Tm value of the primers is designed to be around 60°C. For primers with lower annealing temperatures or for amplifying long fragments exceeding 200 BP, a three-step method is recommended. The extension time can be adjusted based on the length of the PCR product, GC content, and other factors. The extension time per kb of product is closely related to template complexity. Simple templates use 20 秒, normal templates use 30 秒, and complex templates use 1 分分鐘.
2. The PCR reaction conditions should be set differently based on factors such as template, 引子, length of the PCR product, 和GC含量. The final concentration of commonly used primers can be adjusted within the range of 0.2-1.0 μM. The DNA template concentration can also be adjusted based on its concentration. 適用於複雜模板或高 GC 含量, consider extending the denaturation/annealing or extension times, and increasing denaturation/annealing temperature.
3. Use dedicated areas and pipettors before and after amplification, 戴手套, 並經常改變它們. After PCR amplification, do not open the reaction tubes directly. Place them at 4°C or -20°C to cool sufficiently before opening to minimize the risk of PCR product contamination in the experimental environment.