Cleavage of delta and epsilon tubulin PCR products is observed upon PCR purification

Prasad Parab; Jyoti Iyer; Tyler W Johannes

doi:10.17912/micropub.biology.000980

Cleavage of delta and epsilon tubulin PCR products is observed upon PCR purification

MicroPubl Biol. 2023 Oct 10:2023:10.17912/micropub.biology.000980. doi: 10.17912/micropub.biology.000980. eCollection 2023.

Authors

Prasad Parab¹, Jyoti Iyer², Tyler W Johannes³

Affiliations

¹ Chemical Engineering , University of Tulsa, Tulsa, Oklahoma, United States.
² Chemistry and Biochemistry , University of Tulsa, Tulsa, Oklahoma, United States.
³ Chemical Engineering, University of Tulsa, Tulsa, Oklahoma, United States.

Abstract

In this study, we report an unusual phenomenon of the self-cleavage of purified PCR products of codon-optimized Chlamydomonas reinhardtii delta tubulin ( uni3 ) and epsilon tubulin ( bld2 ) genes through an unknown mechanism. Our studies revealed that intact PCR products for both these genes could be obtained upon PCR amplification from plasmid templates carrying these genes. However, interestingly, purification of these PCR products led to their cleavage through an unidentified mechanism. This cleavage persisted despite using different PCR purification kits. Deleting a synthetic intron within the delta tubulin gene also did not have any effect on this cleavage.

Grants and funding

This work was funded by the University of Tulsa Interdisciplinary Project Grant Program awarded to Drs. Jyoti Iyer and Tyler W Johannes. This work was also supported by the University of Tulsa start-up funding to Dr. Jyoti Iyer.