AccuStart II PCR ToughMix

• 2X PCR Master Mix ，加入 DNA 樣本與 Primers 即可進行反應。
  
• ToughMix 專利技術配方，能有效對抗 PCR inhibitors 抑制作用，適合動植物組織、臨床檢體、環境、食物等各種來源 DNA 樣本。
  
• 採用超高純度 Hot-start polymerase ，避免產生非專一性 PCR 產物。
  
• PCR 產物可適用於 TA Cloning 實驗。

Inhibitor Resistance of AccuStart II PCR ToughMix. A 1-kb fragment from 1e4 copies of the Tetracyclin resistance gene was amplified in 20 µl reaction volumes according to the recommended protocol. Reactions were challenged with varying concentrations of different PCR inibitors as summarized below. Following a 3 min activation at 94°C; PCR was for 30 cycles of: 94°C, 15 s; 60°C, 20 s; 72°C, 1 min. 1/5th of each reaction was analyzed on a 01% agarose, 0.5x TBE gel conatining 0.25 mg/ml ethidium bromide. Hemoglobin = 316 ng/µl, 100 ng/µl, 31.6 ng/µl, 10 ng/µl, 3.16 ng/µl Humic Acid = 31.6 ng/µl, 10 ng/µl, 3.16 ng/µl, 1 ng/µl, 0.316 ng/µl Hematin = 100 µM, 31.6 µM, 10 µM, 3.16 µM, 1 µM Melanin = 10 ng/µl, 3.16 ng/µl, 1 ng/µl, 0.316 ng/µl, 0.1 ng/µl C = adenomatous polyposis coli; L = 1 Kb Plus DNA Ladder (Invitrogen) Inhibitor Resistance of AccuStart II PCR ToughMix: PCR in the presence of polyphenol spike. Varying amounts of a polyphenol-rich plant extract (0.2, 0.06, 0.02, 0.006, or 0.002 µl) were added to 25 µl PCRs containing 10,000 copies of a control template. Amplification was carried out for 30 cycles of: 94°C, 15 s; 60°C, 20 s; 72°C, 1 min. 1/5th of each reaction was analyzed on a 01% agarose, 0.5x TBE gel containing 0.25 mg/ml ethidium bromide. As little as 0.002 µl of the crude plant lysate inhibited control reactions with a conventional PCR master mix (data not shown).

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