The Next Step of Cloning

  • Seok-Yong Choi
  • Hyunju Ro
  • Hankuil Yi


When I first started cloning, I feared inserting a DNA fragment into the middle of a gene, such as inserting the EGFP gene into between 96 Leucine (L) and 97 Phenylalanine (F) of Gene A (Fig. 4.1). Insertion here means that EGFP enters into between 96L and 97F without adding any additional nucleotides. When you are extremely lucky (as lucky as winning the lottery), a unique restriction site might fall between 96L and 97F. However, I have never been that lucky. What then can be done? It’s simple. Use PCR. As mentioned before, cloning has become much easier with the advent of PCR.

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Seok-Yong Choi
    • 1
  • Hyunju Ro
    • 2
  • Hankuil Yi
    • 3
  1. 1.Department of Biomedical SciencesChonnam National University Medical SchoolGwangjuSouth Korea
  2. 2.Department of Biological Sciences and BiotechnologyChungnam National UniversityDaejeonSouth Korea
  3. 3.Department of Biological SciencesChungnam National UniversityDaejeonSouth Korea

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