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A Study of the CaCl2 Induced E. Coli DH5-Alpha Transformation by Heat Shock Accompanied by Vibration

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CMBEBIH 2021 (CMBEBIH 2021)

Part of the book series: IFMBE Proceedings ((IFMBE,volume 84))

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Abstract

Chemical methods are commonly used for the induction of Escherichia coli (E. coli) competency to allow transformation mediated by heat shock, an essential procedure for gene cloning. Being the most labor and resource efficient chemical method, the ‘calcium chloride’ (CaCl2) method involves a brief heat shock of bacteria in the presence of Ca2+ ions. Several optimizations were made on this original protocol but the effect of vibration or rotational shaking (revolution per minute) accompanied by the heat shock has not been studied. This paper reports the effect of shaking during or after the heat shock of competent DH5-Alpha strain of E. coli, induced by CaCl2. Our results show that shaking during heat shock does not have any positive impact on the transformation efficiency. On the other hand, after the heat shock, 180 rpm shaking at 37 °C for 1 h has increased transformation efficiency significantly (P < 0.05).

Thus, a post-heat shock incubation with a rotational shaking attains higher yield of transformed cells affecting overall efficiency of transformation of E. coli DH5-Alpha.

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Authors and Affiliations

  1. Genetics and Bioengineering Program, Faculty of Engineering and Natural Sciences, International University of Sarajevo, Hrasnička cesta br. 15, 71210, Sarajevo, Bosnia and Herzegovina

    Lejla-Nur Smajović & Ayla Arslan

  2. Gottfried Schatz Research Center for Cell Signaling, Metabolism and Aging, Medical University of Graz, Neue Stiftingtalstraße 6/6 8010, Graz, Austria

    Furkan Enes Oflaz

  3. School of Advanced Studies, University of Tyumen, Ulitsa 8 Marta, 2/1, Tyumen, 625000, Russia

    Ayla Arslan

Authors
  1. Lejla-Nur Smajović
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  2. Furkan Enes Oflaz
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  3. Ayla Arslan
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Correspondence to Ayla Arslan .

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Editors and Affiliations

  1. Verlab Ltd Sarajevo and Faculty of Pharmacy, University of Sarajevo, Sarajevo, Bosnia and Herzegovina

    Almir Badnjevic

  2. Verlab Ltd Sarajevo and International Burch University Sarajevo, Sarajevo, Bosnia and Herzegovina

    Lejla Gurbeta Pokvić

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Smajović, LN., Oflaz, F.E., Arslan, A. (2021). A Study of the CaCl2 Induced E. Coli DH5-Alpha Transformation by Heat Shock Accompanied by Vibration. In: Badnjevic, A., Gurbeta Pokvić, L. (eds) CMBEBIH 2021. CMBEBIH 2021. IFMBE Proceedings, vol 84. Springer, Cham. https://doi.org/10.1007/978-3-030-73909-6_65

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  • DOI: https://doi.org/10.1007/978-3-030-73909-6_65

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  • Publisher Name: Springer, Cham

  • Print ISBN: 978-3-030-73908-9

  • Online ISBN: 978-3-030-73909-6

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