使用甜菜碱作为共溶剂的新型降落方法，可提高 GC 丰富基因扩增的特异性和强度。

A new approach to touch down method using betaine as co-solvent for increased specificity and intensity of GC rich gene amplification.

机构信息

Department of Endocrinology and Metabolism, Banaras Hindu University, Uttar Pradesh, India.

出版信息

Gene. 2012 Apr 15;497(2):269-72. doi: 10.1016/j.gene.2012.01.031. Epub 2012 Jan 28.

DOI:10.1016/j.gene.2012.01.031

Abstract

Tissue specific genes that contain high GC segments are difficult to amplify by standard PCR. We report an improved method for successful amplification of gene segment that has >70% GC base pairs. This new method of touch down PCR differed by having an initial annealing temperature (Ta) 1.5°C below the primers melting temperature that descended 0.2°C per cycle for 20 cycles and continued thereafter at fixed Ta for next 15 cycles. Different co-solvents were tested with this method to improve the result and betaine proved better than the other co-solvents. This new method is economical, fast and specific in amplifying GC rich region of other genes also.

摘要

组织特异性基因含有高 GC 片段，难以通过标准 PCR 进行扩增。我们报告了一种成功扩增基因片段的改良方法，该基因片段的 GC 碱基对含量> 70%。这种新的降落 PCR 方法的初始退火温度（Ta）比引物解链温度低 1.5°C，每个循环下降 0.2°C，持续 20 个循环，然后在固定 Ta 下继续进行接下来的 15 个循环。用这种方法测试了不同的共溶剂来提高结果，发现甜菜碱比其他共溶剂效果更好。这种新方法经济、快速且特异性地扩增其他基因的 GC 丰富区域。