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[一种用于噬菌体展示肽库中靶分子计数的高效准确方法]

[An efficient and accurate method for counting target molecules in phage-display peptide library].

作者信息

Wang Ke-Geng, Zeng Qing-Ren, Yu Zheng-Yang, Zeng Tie-Bing, Liu Yan

机构信息

Department of Parasitology, School of Medicine, University of South China, Hengyang 421001, China.

出版信息

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2011 Jun;29(3):236-8.

Abstract

The phage titer of samples representing the low, intermediate and high phage number was respectively determined by the double-layer agar plate (DLAP) method and real-time PCR assay. The two methods accurately measured the titer of samples. The plaques from about 1/3 double-agar layer plates could be used to determine the phage titer. The DLAP experiment should repeat 10 times with 10 microl sample each time, while the within-assay coefficient variation (CV) was 4.93%-30.38%. At the same time, the real-time PCR assay only repeated 3 times with 1 microl phage each time, while CV for within-assay ranged from 0.02% to 0.25%. Results indicated that real-time PCR is a simple and quick method for determining bacteriophage titer.

摘要

分别采用双层琼脂平板(DLAP)法和实时荧光定量PCR检测法测定代表低、中、高噬菌体数量的样本的噬菌体效价。这两种方法均能准确测定样本的效价。约1/3双层琼脂平板上的噬菌斑可用于测定噬菌体效价。DLAP实验每次用10微升样本应重复10次,实验内变异系数(CV)为4.93%-30.38%。同时,实时荧光定量PCR检测法每次仅用1微升噬菌体重复3次,实验内CV范围为0.02%-0.25%。结果表明,实时荧光定量PCR是一种测定噬菌体效价的简单快速方法。

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