›› 2011, Vol. 29 ›› Issue (3): 19-236-238.

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An Efficient and Accurate Method for Counting Target Molecules in Phage-Display Peptide Library

 WANG  Ke-Geng, ZENG  Qing-Ren, YU  Zheng-Yang, ZENG  Tie-Bing, LIU  Yan   

  1. 1 Department of Parasitology，School of Medicine，University of South China，Hengyang 421001，China; 2 Cell and Molecular Biological Experiment Center，Xiangya School of Medicine，Central South University，Changsha 410013，China; 3 Department of Surgical Oncology，the First Affiliated Hospital of Unversity of South China，Hengyang 421001，China; 4 Department of Microbiology and Immunology，School of Medicine，University of South China，Hengyang 421001，China
  • Online:2011-06-30 Published:2012-09-27

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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 μl 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 μl 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.

Key words: Phage, Titer, Double layer agar plate method：Real-Time PCR