Cargando…

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

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 b...

Descripción completa

Detalles Bibliográficos
Autores principales: Parab, Prasad, Iyer, Jyoti, Johannes, Tyler W
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Caltech Library 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10594130/
https://www.ncbi.nlm.nih.gov/pubmed/37881246
http://dx.doi.org/10.17912/micropub.biology.000980
_version_ 1785124581097340928
author Parab, Prasad
Iyer, Jyoti
Johannes, Tyler W
author_facet Parab, Prasad
Iyer, Jyoti
Johannes, Tyler W
author_sort Parab, Prasad
collection PubMed
description 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.
format Online
Article
Text
id pubmed-10594130
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Caltech Library
record_format MEDLINE/PubMed
spelling pubmed-105941302023-10-25 Cleavage of delta and epsilon tubulin PCR products is observed upon PCR purification Parab, Prasad Iyer, Jyoti Johannes, Tyler W MicroPubl Biol New Finding 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. Caltech Library 2023-10-10 /pmc/articles/PMC10594130/ /pubmed/37881246 http://dx.doi.org/10.17912/micropub.biology.000980 Text en Copyright: © 2023 by the authors https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle New Finding
Parab, Prasad
Iyer, Jyoti
Johannes, Tyler W
Cleavage of delta and epsilon tubulin PCR products is observed upon PCR purification
title Cleavage of delta and epsilon tubulin PCR products is observed upon PCR purification
title_full Cleavage of delta and epsilon tubulin PCR products is observed upon PCR purification
title_fullStr Cleavage of delta and epsilon tubulin PCR products is observed upon PCR purification
title_full_unstemmed Cleavage of delta and epsilon tubulin PCR products is observed upon PCR purification
title_short Cleavage of delta and epsilon tubulin PCR products is observed upon PCR purification
title_sort cleavage of delta and epsilon tubulin pcr products is observed upon pcr purification
topic New Finding
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10594130/
https://www.ncbi.nlm.nih.gov/pubmed/37881246
http://dx.doi.org/10.17912/micropub.biology.000980
work_keys_str_mv AT parabprasad cleavageofdeltaandepsilontubulinpcrproductsisobserveduponpcrpurification
AT iyerjyoti cleavageofdeltaandepsilontubulinpcrproductsisobserveduponpcrpurification
AT johannestylerw cleavageofdeltaandepsilontubulinpcrproductsisobserveduponpcrpurification