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Homemade 100 Pair Base DNA Ladder Based on Bacterial Plasmid pMAL-C2X

Firuzeh Badreh1, Khodakaram Jahanbin2, Ali Khodadadi2, Ali Khorasani Zadeh2, Moosa Sharifat2, Milad Khayati2, Mohammad Rashno2,3

Background: Most molecular biology research findings require markers such as DNA ladders to detect and quantify nucleic acid fragments length by electrophoresis in routine task.

Methods: In this study, we report a method to prepare a 100 pair base DNA ladder based on PCR technique. The bacterial plasmid pMAL-C2X was used as the template DNA in PCR. PCR products were extracted by phenol/chloroform, precipitated with ethanol and mixed proportionally.

Results: Finally, a set of 7 primers (5 revers and 2 forward) successfully designed. The amplified PCR fragments were quantified by nanodrop and the bands size were estimated by gel electrophoresis along with a co-migrating commercial 100 bp ladder in 1.5% agarose gel. The results indicated that the clear and sharp bands of 100 bp-1000 bp were successfully generated by one reaction in PCR.

Conclusion: Our homemade product is a competitive product with a commercial market and can be prepared with a simple and inexpensive method and low probability of non-specific band.