2× Taq 聚合酶鍊式反應 Master Mix（with dye）
編號： E002 規格：1mL Storage: 儲存於-20°C
產品介紹：
The 2x Taq PCR Master Mix (帶染料) is a premixed solution at twice the concentration of DNA Polymerase, PCR緩衝液, dNTP Mix, and indicators. When using this product, simply add the template and primers to the premixed solution, and complete the volume with ddH2O for the PCR reaction. This simplifies the operational steps and reduces the risk of contamination during PCR procedures. 另外, the premixed solution contains the necessary dye reagents for gel electrophoresis, enabling direct gel electrophoresis after PCR. This product offers high sensitivity, strong specificity, and a wide linear range, allowing for more accurate quantification of target genes. The PCR products amplified using this product have an added A base at the 3′ 結尾, facilitating cloning into T vectors.
產品內容：
| 成分 | E002 |
| 2× Taq PCR Master Mix（with dye） | 1毫升 |
貯存:
儲存於-20°C, 最短保存期限為 12 月.
品質管制:
本產品經過品質檢測，不含核酸內切酶活性, 核酸外切酶活性, 和核糖核酸酶污染. 殘留宿主基因組DNA如下 10 副本.
產品用途:
使用PCR方法擴增DNA; DNA序列測定.
使用說明:
- Allow all required PCR reaction solutions to equilibrate to room temperature until fully dissolved. Thoroughly mix the solutions and set up the PCR reaction system on ice.
- It is recommended to set up the PCR reaction system on an ice bath or in a cooler, 以下推薦反應體系供參考.
- 用移液管將準備好的反應體系徹底吹打並混合, 用蓋子密封 PCR 管, 給它們貼上適當的標籤, 並在室溫下短暫離心.
- Place the prepared PCR tubes into the PCR機, 設定反應條件, 並啟動PCR反應.
推薦反應系統:
| 試劑 | 25µL 系統體積 | 最終濃度 |
| 2× Taq PCR Master Mix（with dye） | 12.5微升 | 1× |
| 底漆一 (10微米) | 0.5-2.5微升 | 0.2-1.0μm |
| 第一二號 (10微米) | 0.5-2.5微升 | 0.2-1.0μm |
| 模板DNA | 1μL as required | — |
| DDH2氧 | Up to 25μL | — |
反應條件:
Under normal circumstances, a two-step method can be used for the reaction. If the two-step amplification is not satisfactory, a three-step method can be employed to set up the PCR reaction program. Considering the uncertainty of target gene abundance in cDNA templates, it is recommended to use 35-38 cycles for the initial amplification and adjust the number of cycles based on the amplification results.
| 方法/步驟 | 時間設定 | 週期 |
| 95℃ (預變性) | 2-5分分鐘 | 1 |
| 95℃ (變性) | 10秒 | 28-40週期
（Plasmid or Genomic Template：28-35週期； cDNA Template：30-40cycles） |
| 55℃-60℃ (退火) | 30秒 | |
| 72℃ (擴大) | 1分分鐘 / 1-3知識庫 | |
| 72℃ (最終延期) | 5-10分分鐘 | 1 |
| 4℃/16℃（Hold） | 無窮大 | — |
筆記: 反應條件可依實際要求進行調整和最佳化.
預防措施:
- 延伸時間可依PCR產物的長度、GC含量等因素調整. 產品每kb的延伸時間與模板的複雜程度密切相關: 簡單的模板需要 20 秒, 典型的模板需要 30 秒, 並且需要複雜的模板 1 分分鐘.
- PCR 反應的設定應根據不同的條件（例如模板）進行定制, 引子, PCR產物長度, 和GC含量. The final concentration of primers typically falls within the range of 0.2-1.0μM. DNA模板濃度可隨之調整. 適用於複雜模板或高 GC 含量, 建議延長預變性/變性或延伸時間並提高變性/退火溫度.
- 擴增前後使用指定區域和移液器, 戴手套, 並經常改變它們. PCR反應完成後, 不要立即打開反應管. 打開前讓其在 4°C 或 -20°C 下充分冷卻，以最大程度地降低 PCR 產品污染實驗室環境的風險.
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