计算溶液所需的质量、体积或浓度。
,适用于对基线干扰要求严格的色谱和分析工作流程。
-20°C储存,避免反复冻融。超低温运输 。请查阅批次 COA 获取详细规格。
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在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
产品内容
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产品简介
HiFi PCR Mix for NGS是由热启动酶、PCR Buffer、dNTPs、Mg2+以及PCR稳定剂和增强剂等组成的预混体系 , 具有高保真性、高延伸性、低偏好性的特点 , 对复杂的DNA模板(如高GC含量模板)有均衡的扩增效率 , 特别适合千二代建库中多重PCR的扩增。
本品所含的高效热启动酶,在常温下没有聚合酶活性,有效避免了在常温条件下由引物和模板非特异性结合或引物二聚体而产生的非特异性扩增 。独特的缓冲体系与热启动酶的组合, 显著提高了PCR的扩增效率, 扩增范围更广泛。本产品有效提升了基因组中高GC或高AT区域的扩增效率, 降低扩增偏好性从而提高测序覆盖度。
注意事项
1. 该产品不适用千有修饰的引物 。
2. 使用前请上下颠倒轻轻混匀, 尽量避免起泡, 并经短暂离心后使用。
3. . 避免反复冻融, 反复冻融可能使产品性能下降。如果在短期内需要频繁使用, 可在2-8°C 保存。
使用方法
以下举例为常规P CR 反应体系和反应条件 , 实际操作中应根据模板、引物结构和目的片段大小不同进行相应的改进和优化。
1. PCR 反应体系
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注意: 引物浓度请以终浓度0.1-0.3µM作为设定范围的参考。
2. PCR 反应条件 |

注意:
1) 一般实验中退火温度比扩增引物的熔解温度 Tm低5°C , 无法得到理想的扩增效率时, 适当降低退火温度;发生非特异性反应时,提高退火温度,巾此优化反应条件。
2 ) 可根据扩增产物的下游应用设定循环数。如果循环次数太少 , 扩增量不足; 如果循环次数太多, 错配机率会增加, 非特异性背景严重。所以在保证产物得率的前提下应尽量减少循环次数。
Product content
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Product Introduction
HiFi PCR Mix for NGS is a premixed system consisting of hot starter enzyme, PCR Buffer, dNTPs, Mg2+, and PCR stabilizers and enhancers, etc. It is characterized by high fidelity, high elongation, and low preference, and has a balanced amplification efficiency for complex DNA templates (e.g., high GC-content templates), and it is especially suitable for multiplexed PCR in the thousand-two library building .
The high efficiency hot starter enzyme contained in this product has no polymerase activity at room temperature, which effectively avoids non-specific amplification caused by non-specific binding of primers and templates or primer dimerization under room temperature conditions. The unique combination of buffer system and hot starter enzyme significantly improves the amplification efficiency of PCR with wider amplification range. This product effectively enhances the amplification efficiency of high GC or high AT regions in the genome, reduces amplification preference and thus improves sequencing coverage.
matters needing attention
This product is not suitable for thousands of primers with modifications.
Before use, please mix gently by turning up and down, avoid foaming as much as possible, and use after brief centrifugation. Avoid repeated freezing and thawing, which may deteriorate the performance of the product. If frequent use is required within a short period of time, store at 2-8°C.
Usage
The following examples are conventional PCR reaction systems and reaction conditions, which should be improved and optimized according to the template, primer structure and target fragment size in actual operation.
PCR reaction system
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Note: Please use the final concentration of 0.1-0.3µM as a reference for setting the range of primer concentration.
2. PCR reaction conditions
Attention:
In general, the annealing temperature was 5°C lower than the melting temperature Tm of the amplification primers, and when the desired amplification efficiency could not be obtained, the annealing temperature was lowered appropriately; when non-specific reactions occurred, the annealing temperature was increased, and the reaction conditions were optimized.
2) The number of cycles can be set according to the downstream application of the amplification product. If the number of cycles is too low, the amplification is insufficient; if the number of cycles is too high, the chance of mismatch will increase and the non-specific background will be serious. Therefore, the number of cycles should be minimized under the premise of ensuring the product yield.