Main Article Content
DNA ladder has been widely used to determine the size of DNA fragments by electrophoresis in routine molecular biology laboratories. Commercial DNA ladders are typically used for comparing the size and estimating the DNA concentration of unknown DNA sample. However, it is quite expensive. Here, we developed a simple strategy for the preparation of 100 bp DNA Ladder which comprises the 12 designed primers to amplify 100 bp to 1,000 bp and 1,500 bp DNA fragments from the vector (pRGEB32) as a template for polymerase chain reaction (PCR). All primers could be used for amplification of all individual fragments in the same PCR profile. Therefore, the total running time of PCR was reduced. In addition, there were no undesired PCR products and the requirement of the purification step. Our procedure for production of the DNA ladder was simple and time saving. The total cost was also inexpensive compared with those of the commercial ones. This result indicates that it can be used for molecular studies.
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
บทความที่ได้รับการตีพิมพ์เป็นลิขสิทธิ์ของ PBRU Science Journal
Polyarush SV, Egamberdiev SS, Mansurov DR, Azimova SS. Preparation of DNA markers based on E. coli plasmid DNA. Chemistry of Natural Compounds 2003;39:592-4.
Henrici RC, Pecen TJ, Johnston JL, Tan S. The pPSU Plasmids for Generating DNA Molecular Weight Markers. Scientific Reports 2017;1(2438). Doi:10.1038/s41598-017-02693-1.
Parker RC, Watson RM, Vinograd J. Mapping of closed circular DNAs by cleavage with restriction endonucleases and calibration by agarose gel electrophoresis. Proceedings of the National Academy of Sciences of the United States of America 1977;74(3):851-5.
Cooney CA. Techniques and high-resolution DNA size markers for pulsed field gel electrophoresis. Molecular Biotechnology 1994;2:119-27.
Abbasian M, Seyedi HA, Boroujeni ZK, Mofid MR. Easy method for production of a home-made DNA ladder in every laboratory. Advanced Biomedical Research 2015;4(70). Doi:10.4103/2277-9175.153894.
Xie K, Minkenberg B, Yang Y. Boosting CRISPR/Cas9 multiplex editing capability with the endogenous tRNA-processing system. Proceedings of the National Academy of Sciences of the United States of America 2015;112(11): 3570-5.
Wu J, Ye C. Tandem PCR: A novel and efficient unit amplification model for the preparation of small DNA fragments. Molecular Biology Reports 2011;38:2729-31.
Amills M, Francino O, Sánchez A. Primer-directed synthesis of a molecular weight marker. Genetic analysis 1996;13:147-9.
Chang M, Wang JH, Lee HJ. Laboratory production of 100 base pair DNA molecular weight markers. Journal of Biochemical and Biophysical Methods 2008;70:1199-1202.
Wang TY, Guo L, Zhang JH. Preparation of DNA ladder based on multiplex PCR technique. Journal of Nucleic Acids 2010;421803. Doi:10.4061/2010/421803.
Rashno M, Shapouri MRSA, Jolodar A. Construction of a synthetic vector for preparation of a 100 base pair DNA ladder. Iran Journal of Biotechnology 2012;10:106-10.
Lan VTT, Loan PTT, Duong PAT, Thanh LT, Ha NT, Thuan TB. Straightforward Procedure for Laboratory Production of DNA Ladder. Journal of Nucleic Acids 2012; Doi:10.1155/2012/254630.