COLD-PCR (co-amplification at lower denaturation temperature PCR) is a modified polymerase chain reaction (PCR) protocol that enriches variant alleles from a mixture of wildtype and mutation-containing DNA.

COLD-PCR can be applied in the place of conventional PCR, and analyzed either in real time or via most downstream molecular applications. COLD-PCR currently has three platforms: full-COLD-PCR, fast-COLD-PCR and ice-COLD-PCR. An appropriate platform can be utilized depending upon the desired level of enrichment and the type of variant(s) to be ...

2022年2月10日 · 低温变性共扩增 PCR，是不管突变存在于什么位置，均能从野生型和含突变的序列的基因混合物中选择性扩增出为数不多的等位基因的方法。 目前，用于低温变性共扩增 PCR 的方法主要有 1 种：低温变性共扩增 PCR。 低温变性共扩增 PCR 的基本原理是，双链 DNA 合成时，由于单个核苷酸的错配，使得这段序列的熔点温度会产生微小的而可预测的变化。 根据这段序列的上下游碱基背景和错配位置，多至 200 bp 的序列的熔点温度一般可以变化 0.2~1.5 ℃ 甚至 …

Co-amplification at lower denaturation temperature-based polymerase chain reaction (COLD-PCR) is a single-step amplification method that results in the enhancement of both known and unknown minority alleles during PCR, irrespective of mutation type and position. This method is based on exploitation …

2018年8月31日 · 近期出现了低温变性共扩增PCR技术( co-amplification at lower denaturation temperature- PCR, COLD-PCR),这种新型的PCR方法,不管突变存在于什么位置,均能从野生型和含突变的序列的基因混合物中选择性扩增出为数不多的等位基因。因此, COLD-PCR从基因组中扩增出发生突变的等位 ...

2016年2月4日 · Co-amplification at lower denaturation temperature-based polymerase chain reaction (COLD-PCR) is a single-step amplification method that results in the enhancement of both known and unknown minority alleles during PCR, irrespective of mutation type and position.

2023年8月22日 · COLD-PCR依赖的理论是杂合DNA双链的碱基不匹配，从而导致双链结合稳定性下降，即杂合DNA双链的变性温度降低。 于是，在以野生型和含有突变的序列的基因混合物 …

Co-amplification at lower denaturation temperature-based polymerase chain reaction (COLD-PCR) is a single-step amplification method that results in the enhancement of both known and unknown minority alleles during PCR, irrespective of mutation type

一、基本原理：PCR技术的基本原理类似于DNA的天然复制过程，其特异性依赖于与靶序列两端互补的寡核苷酸引物。 DNA的半保留复制是生物进化和传代的重要途径。 双链DNA在多种酶的作用下可以变性解旋成单链，在DNA聚合酶的参与下，根据碱基互补配对原则复制成同样的两分子拷贝。 在实验中发现，DNA在高温时也可以发生变性解链，当温度降低后又可以复性成为双链。 因此，通过温度变化控制DNA的变性和复性，加入设计引物，DNA聚合酶、dNTP就可以完成特定 …

COLD-PCR is based on the observations that (a) for each DNA sequence there is a critical denaturation temperature (Tc) that is lower than the Tm of the target sequence and below which PCR efficiency drops abruptly, and (b) Tc is dependent on the DNA sequence.