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AAT Bioquest

What are the main steps of cloning?

Posted June 22, 2020


Answer

In standard molecular cloning experiments, the cloning of DNA fragment involves seven main steps:

  1. Isolation of DNA (gene of interest) fragments to be cloned
  2. Insertion of isolated DNA into a suitable vector to from the recombinant DNA
  3. Introduction of the recombinant DNA into a suitable host organism
  4. Selection of the transformed host cells and identification of the clone containing the gene of interest
  5. Multiplication/expression of the introduced gene in the host
  6. Isolation of the multiplied gene copies/protein expressed by the intro­duced gene
  7. Purification of the isolated gene copy/protein
Additional resources

Helixyte™ Green *10,000X Aqueous PCR Solution*

6-ROX glycine *25 uM fluorescence reference solution for PCR reactions*

Perdew, G. H., Heuvel, J. P. V., & Peters, J. M. (2008). Regulation of gene expression. Springer Science & Business Media.

Lodish, H., Berk, A., Kaiser, C. A., Krieger, M., Scott, M. P., Bretscher, A., ... & Matsudaira, P. (2008). Molecular cell biology. Macmillan.