Zymo Research has simplified my life with their DNA Clean & Concentrator-5 kit. Recently, I tried this kit for the first time and I was truly amazed at how fast and easy it is to use. This kit is primarily for concentrating DNA (up to 5 ug) that has been generated in a PCR reaction or to clean DNA that has been used in a restriction enzyme (or phosphatase/kinase) reaction. The beauty of the system is the minimal time required to isolate the DNA as well as the small elution volumes. Zymo Research states that the DNA can be isolated in 2 minutes, but realistically it’s 5 minutes from start to finish, with the DNA eluted in a minimal volume of 6 ul.
The DNA Clean & Concentrator is based on Zymo Research’s single buffer procedure and Zymo-spin columns. The DNA to be concentrated is re-suspended in 2 volumes of the DNA Binding Buffer and passed through the spin column. The bound DNA is washed twice with the supplied wash buffer and finally eluted with water or 10 mM Tris-HCl, 1mM EDTA, pH 8.0.
Purifying high quality DNA in a minimal volume saved time by eliminating ethanol precipitation and desalting (by drop dialysis) steps. Based on ethidium bromide staining of DNA standards and dilutions of the purified sample, DNA recovery was close to 90% with the NEB pBeloBAC11 vector. However, plasmid recovery of 12-kbp plasmid was closer to 50%.
This kit is becoming a staple in our lab. The time saved in down-stream applications and its reasonable cost ($62.00 for 50 samples) make this kit a great buy. Our lab will probably never use another method for cleaning and concentrating DNA.
Vincent Magrini, Ph.D.
Department of Molecular Microbiology
Washington University School of Medicine