Optimization of mutation detection methods in the NUD gene induced by CRISPR / CAS9 technology in barley (Hordeum vulgare L.)
Mateusz Przyborowski
m.przyborowski@ihar.edu.plZakład Genomiki Funkcjonalnej,2 Zakład inżynierii Genetycznej Instytut Hodowli i Aklimatyzacji Roślin — Państwowy Instytut Badawczy, Radzików (Poland)
https://orcid.org/0000-0001-7456-3808
Sebastian Gasparis
Zakład Genomiki Funkcjonalnej,2 Zakład inżynierii Genetycznej Instytut Hodowli i Aklimatyzacji Roślin — Państwowy Instytut Badawczy, Radzików (Poland)
https://orcid.org/0000-0003-0521-086X
Wacław Orczyk
Zakład Genomiki Funkcjonalnej,2 Zakład inżynierii Genetycznej Instytut Hodowli i Aklimatyzacji Roślin — Państwowy Instytut Badawczy, Radzików (Poland)
https://orcid.org/0000-0003-4735-0000
Anna Nadolska-Orczyk
Zakład Genomiki Funkcjonalnej,2 Zakład inżynierii Genetycznej Instytut Hodowli i Aklimatyzacji Roślin — Państwowy Instytut Badawczy, Radzików (Poland)
https://orcid.org/0000-0001-6127-3860
Abstract
The aim of the study was to select the optimal method for mutation detection in the Nud gene induced by using the CRISPR / Cas9 technology in barley. Four commonly used genotyping techniques were tested: restriction fragment length polymorphism (RFLP), enzymatic digestion using endonuclease I from T7 bacteriophage, high resolution melting curve of polymerase chain reaction product (HRM-PCR) and molecular probes. Findings in the current study suggest that the most favorable way for detecting mutations induced by the CRISPR / Cas9 technique is a method based on molecular probes. It enables to receive clear and repeatable results but at the same time requires the use of advanced equipment.
Keywords:
CRISPR / Cas9, genome editing, Nud gene, barley, molecular probesPrzyborowski, M. (2019) “Optimization of mutation detection methods in the NUD gene induced by CRISPR / CAS9 technology in barley (Hordeum vulgare L.)”, Bulletin of Plant Breeding and Acclimatization Institute, (287), pp. 33–34. doi: 10.37317/biul-2019-0104.
Authors
Mateusz Przyborowskim.przyborowski@ihar.edu.pl
Zakład Genomiki Funkcjonalnej,2 Zakład inżynierii Genetycznej Instytut Hodowli i Aklimatyzacji Roślin — Państwowy Instytut Badawczy, Radzików Poland
https://orcid.org/0000-0001-7456-3808
Authors
Sebastian GasparisZakład Genomiki Funkcjonalnej,2 Zakład inżynierii Genetycznej Instytut Hodowli i Aklimatyzacji Roślin — Państwowy Instytut Badawczy, Radzików Poland
https://orcid.org/0000-0003-0521-086X
Authors
Wacław OrczykZakład Genomiki Funkcjonalnej,2 Zakład inżynierii Genetycznej Instytut Hodowli i Aklimatyzacji Roślin — Państwowy Instytut Badawczy, Radzików Poland
https://orcid.org/0000-0003-4735-0000
Authors
Anna Nadolska-OrczykZakład Genomiki Funkcjonalnej,2 Zakład inżynierii Genetycznej Instytut Hodowli i Aklimatyzacji Roślin — Państwowy Instytut Badawczy, Radzików Poland
https://orcid.org/0000-0001-6127-3860
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