Creating Random Mutagenesis Libraries by Megaprimer PCR of Whole Plasmid (MEGAWHOP)

  • Kentaro Miyazaki
Part of the Methods in Molecular Biology™ book series (MIMB, volume 231)


The conventional method for cloning a DNA fragment is to insert it into a vector and ligate it (1). Although this method is commonly used, it is labor intensive because the ratio and concentrations of the DNA insert and the vector must be optimized. Even then, the resulting library is often plagued with unwanted plasmids that have no inserts or multiple inserts. These species have to be eradicated to avoid tedious screening, especially when producing random mutagenesis libraries.


Exonuclease Activity Template Plasmid Specific Restriction Enzyme Single Insert Multiple Insert 
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Copyright information

© Humana Press Inc., Totowa, NJ 2003

Authors and Affiliations

  • Kentaro Miyazaki
    • 1
  1. 1.National Institute of Advanced Industrial Science and Technology (AIST)IbarakiJapan

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