Setting up the DNA ladder Production line in order to use in electrophoresis

Sahandi Khalifeh-Kandy, Amin; Pazhouhandeh, Maghsoud

doi:10.30473/cb.2025.72897.1991

Setting up the DNA ladder Production line in order to use in electrophoresis

Document Type : Research Paper

Authors

¹ Biotechnology Dept. Agriculture Faculty, Azarbaijan Shahid Madani University, Tabriz, IRAN

² Biotechnology Department, Agriculture Faculty, Azarbaijan Shahid Madani University, Km 35 Tabriz-Azarshahr Road, Tabriz, Iran.

10.30473/cb.2025.72897.1991

Abstract

DNA markers are the solution of DNA fragments of different lengths. These markers are used to evaluate the results of the polymerase chain reactions (PCR) and the enzymatic digestion reactions, to confirm the vectors containing the desired DNA fragment, to study the genetic variation by molecular markers in different organisms and in other methods used in molecular genetics, biotechnology and in medical diagnostic applications. DNA ladder is an essential tool in agarose or polyacrylamide gel electrophoresis and its price is high due to the high production cost. By comparing the movement of known fragments of a ladder on the electrophoresis gel, the size (base pair) of unknown fragments obtained in a test can be estimated. In this study a new method for producing a high-consumption and low-production-cost DNA size marker was presented. The PCR technique was performed on a plant gene to obtain the desired fragments with different sizes of 100 bp. The interested fragments were cloned separately into pKS plasmid using an enzymatic digestion site at both ends. The recombinant plasmids were amplified in bacteria. Finally, by a simple enzymatic digestion with a common enzyme (BamHI) on plasmids extracted from bacteria, all the necessary fragments were obtained to produce the expected DNA size marker. In this applicable research, the production line of a DNA marker was set up. This produced ladder with a range of 100 to 3000 bp fragments has a wide use in medical research and diagnosis.

Keywords

Main Subjects

Molecular Genetics and Genetic Engineering

References

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