Taq DNA Polymerase
商品番号:E001B 仕様:500U/1000U/5000U Storage: -20℃で保存
製品導入:
This product is Taq DNA Polymerase, commonly referred to as Taq enzyme, which is one of the most widely used DNA polymerases. It is expressed in Escherichia coli with the cloned gene of Thermus aquaticus DNA Polymerase and then purified through multiple steps. The PCR products amplified using this product have an added A base at the 3′ end, allowing for cloning into T vectors.
Product Features:
Sanshibio Taq DNA Polymerase is an improved and screened Taq DNA polymerase. In addition to the characteristics of the traditional enzyme itself, it also has broader advantages after repeated testing.
First, Sanshibio’s Taq enzyme is equipped with a basic universal buffer, which has a good amplification effect on common plant, animal, microbial, 等. DNA or cDNA; it is compatible with the buffers of most mainstream manufacturers on the market; the modified The enzyme combined with the basic universal buffer can achieve stronger resistance, and can tolerate DNA crude extracts containing a large amount of impurities; it has stronger stability and can tolerate multiple freeze-thaw cycles; for some low-abundance templates, it can be used The amount of enzyme added is reduced, and the amount of magnesium ions is appropriately increased to achieve more sensitive detection.
商品内容:
コンポーネント | E001B-01(500U) | E001B-02(1000U) | E001B-03(5000U) |
Taq DNA Polymerase(5U/µL) | 100μL | 200μL | 1mL |
dNTP Mixture (10mM each) | 100μL | 200μL | 1mL |
10× PCR Buffer (マグネシウム2+ free)
| 1mL | 2× 1mL | 10× 1mL |
MgCl2 (25mM) | 1mL | 2× 1mL | 10× 1mL |
ストレージ:
-20℃で保存, 最低保存期間は 12 月.
アクティビティの定義:
Using activated largemouth bass sperm DNA as a template/primer, the intake of 10 nmol of total nucleotides as acid-insoluble material within 30 minutes at 74°C is defined as 1 unit of activity (U).
品質管理:
This product has undergone quality testing and is free from endonuclease activity, exonuclease activity, リボヌクレアーゼの汚染. Residual host genomic DNA is below 10 コピー.
Product Usage:
Amplification of DNA using PCR method; DNA sequence determination.
Usage Instructions:
- Dissolve and mix all required solutions for the PCR reaction and place them on an ice bath or in a cooler. It is recommended to aliquot the PCR reaction mixture to avoid repeated freeze-thaw cycles.
- It is advisable to set up the PCR reaction system on an ice bath or in a cooler, following the recommended reaction system for reference.
- Excessive template DNA can lead to nonspecific PCR products. Recommended template amounts for different types of templates in a 50µL reaction volume are as follows:
Genomic DNA from Animals and Plants: 0.1-1μg;
Genomic DNA from Escherichia coli: 10-100の;
プラスミドDNA: 0.1-10の.
Recommended Reaction System:
試薬 | 50µL System Volume | 最終濃度 |
Taq DNA Polymerase | 1μL | 0.1U |
10× PCR Buffer (マグネシウム2+ free) | 5μL | 1× |
MgCl2 (25mM) | 4μL | 2mM |
dNTP Mixture (10mM each) | 1μL | 0.2mM each |
プライマーI (10μM) | 1μL | 0.2μM |
ファーストⅡ (10μM) | 1μL | 0.2μM |
テンプレートDNA | 1μL | — |
ああ2○ | To 50µL | — |
注記: The amounts of each component in the reaction system can be adjusted according to actual requirements.
- Thoroughly pipette and mix the prepared reaction system using a pipette, seal the PCR tubes with caps, label them appropriately, and briefly centrifuge at room temperature.
- Place the prepared PCR tubes into the PCR装置, set the reaction conditions, and initiate the PCR reaction.
Reaction Conditions:
方法・手順 | Time setting | サイクル |
95℃ (変性前) | 2-5分 | 1 |
95℃ (変性) | 30秒 | 25-35 サイクル |
55℃-60℃ (アニーリング) | 30秒 | |
72℃ (拡大) | 1分 | |
72℃ (Final Extension) | 10分 | 1 |
4℃(Hold) | ∞ | — |
注記: Reaction conditions can be adjusted and optimized according to actual requirements.
予防:
- Extension time can be adjusted based on factors such as the length and GC content of the PCR product. The extension time per kb of the product is closely related to the complexity of the template: simple templates require 20 秒, typical templates require 30 秒, and complex templates require 1 分.
- The setup of the PCR reaction should be tailored to different conditions such as template, primers, length of PCR product, and GC content. The final concentration of primers typically falls within the range of 0.1-1.0μM. DNA template concentration can be adjusted accordingly. For complex templates or high GC content, it’s recommended to prolong pre-denaturation/denaturation or extension times and increase denaturation/annealing temperatures.
- Use designated areas and pipettes before and after amplification, 手袋を着用する, そして頻繁に変更する. PCR反応終了後, do not open the reaction tubes immediately. Allow them to cool sufficiently at 4°C or -20°C before opening to minimize the risk of PCR product contamination to the laboratory environment.
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