| Product Description | Aladdin's UltraBio™ PCR SuperMix (Green, 2X) is a 2X concentrated solution of Taq DNA Polymerase, dNTPs, and all of the components required for PCR, except DNA template and primers. It also contains 2X loading buffer that enables PCR products to be directly loaded on an agarose gel for electrophoresis. This product is designed for DNA amplification, qualitative or semi-quantitative PCR analysis, and can also be used for amplification and cloning of DNA fragments less than 2kb.This pre-mixed formulation saves time and avoids contamination by reducing the number of pipetting steps required for PCR, enabling the consistency and the reproducibility of PCR reactions.The UltraBio™ PCR SuperMix (Green, 2X) can be used to amplify DNA fragments up to 5kb in length, but is more suitable for amplification of DNA fragments less than 2kb.Blue and orange dyes (overall green) are added to this product and migrate at approximately 4kb and 50bp, respectively, in 1% agarose gel, which has no effect on the observation of PCR products. PCR products can be loaded directly onto an agarose gel without addition of loading buffers.This product employs a new Taq DNA Polymerase with highly stable enzyme activity. Repeated freeze-thaw up to 15 cycles does not cause any obvious change of its activity. Please refer to Figure 1 for the PCR products amplified using this product with or without 15 freeze-thaw cycles. Different primer sets are used to amplify the DNA fragments in different size ranging from 100bp to 1500bp.Figure 1. Agarose gel electrophoresis of the PCR products amplified using Aladdin's UltraBio™ PCR SuperMix (Green, 2X) without freeze-thaw (A) or with 15 cycles of freeze-thaw (B). All other conditions used for A and B are exactly the same.The PCR product amplified using this product has sticky termina with 3'-dA overhangs, which can be directly ligated to T vector for TA cloning.Each ml of this product is sufficient for 40 PCR reactions of 50μl or 100 PCR reactions of 20μl.
Precautions： Because PCR reactions are extremely sensitive, contamination must be avoided during the preparation of PCR reactions. Negative control without templates is recommended for all PCR assays.The error probability of each cycle of the new Taq DNA polymerase in this product is about 2×10-5. For cloning of DNA fragments larger than 1kb, we recommend using DNA polymerases with lower error probability, such as DNA Polymerase , Plus DNA Polymerase , Q6U™ High-Fidelity DNA Polymerase , Amp™ Extra-long DNA Polymerase or Amp™ Plus Extra-long DNA Polymerase , etc. For general PCR or RT-PCR, this product is an ideal choice.Although repeated freeze-thaws up to 15 cycles do not cause any obvious change of the performance of this product, it should still be avoided. Too many cycles of freeze-thaw can decrease the activity of this product.Thaw this product completely prior to use. Mix well gently by inversion the tube and avoid the formation of air bubbles.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear lab coat and disposable gloves during the operation.
Instructions for Use： 1. Prepare PCR reaction mixture: a. Thaw the Easy-Load™ PCR Supermix (Green, 2X) at room temperature. Mix well gently by inverting the tube and centrifuge briefly at a low speed.b. Set up the PCR reaction on ice as follows:ReagentFinal ConcentrationVolumeVolumeNuclease-Free Water-(21-x)µl(8.4-y)µlTemplate DNA10pg-1µgxµlyµlPrimer Mix (10µM)0.8µM4µl1.6µlEasy-Load™ PCR SuperMix (Green, 2X)1X25µl10µlTotal Volume-50µl20µlNote: (a) Primers at a final concentration of 0.2μM each normally work well, but primer concentration can be optimized between 0.1-1.0μM. Increase the primer concentration when amplification efficiency is low and decrease the primer concentration when non-specific PCR products are amplified. (b) The recommended amount of template varies for different types of DNA. In a reaction volume of 50μl, use 100 ng of Mammalian genomic DNA, 100 ng of E. coli genomic DNA, or 5-30 ng of Plasmid DNA. Too much template DNA can easily lead to non-specific PCR amplification products.a. Mix well gently by vortex or pipetting. Centrifuge briefly at room temperature to collect liquid at the bottom of PCR tube.b. (Optional) When using a thermal cycler without a heated lid, place a drop of mineral oil ( Plus Extra-long DNA Polymerase1000UD7215S Plus Extra-long DNA Polymerase200UD7216Pfu DNA Polymerase100UD7217Pfu DNA Polymerase1000UD7218 High-Fidelity DNA Polymerase2000UD7239MQ6U™ High-Fidelity DNA Polymerase500UD7239SQ6U™ High-Fidelity DNA Polymerase100UD7255-100mlEasy-Load™ PCR Master Mix (Green, 2X)10000TD7255-1mlEasy-Load™ PCR Master Mix (Green, 2X)100TD7255-20mlEasy-Load™ PCR Master Mix (Green, 2X)2000TD7255-4mlEasy-Load™ PCR Master Mix (Green, 2X)400TD7256-1mlEasy-Load™ PCR SuperMix (Green, 2X)100TD7256-20mlEasy-Load™ PCR SuperMix (Green, 2X)2000TD7256-4mlEasy-Load™ PCR SuperMix (Green, 2X)400TD7259-100mlEasy-Load™ PCR Master Mix (Orange, 2X)10000TD7259-1mlEasy-Load™ PCR Master Mix (Orange, 2X)100TD7259-20mlEasy-Load™ PCR Master Mix (Orange, 2X)2000TD7259-4mlEasy-Load™ PCR Master Mix (Orange, 2X)400T
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