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. 2021 Jun 22;9(7):1351. doi: 10.3390/microorganisms9071351

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Standardization of CRISPR_loci-PCR approach using designed F1–R1 primer set. (a) Polymerases (P: GoTaq^TM Flexi from Promega Inc.; K: KAPA Taq PCR Kit from KAPA Biosystems; I: Invitrogen^TM Taq DNA Polymerase from ThermoFisher Scientific Inc.). (b) Primer concentrations (µM). (c) dNTPs concentrations (mM); (d) Dimethyl sulfoxide (DMSO) percentage. Electrophoresis on (e) Tris-Borate-EDTA (TBE) and (f) Sodium borate (SB) agarose gels. L: 100 bp DNA ladder; NC: negative control; +: positive control (pure DNA from Azospirillum sp. strain B510).