DNA Clean and Concentrator 25™ From Zymo Research

Zymo Research’s DNA Clean & Concentrator-25™ product is a hassle-free method for the rapid purification and concentration of up to 25 μg of high quality DNA from PCR, endonuclease digestions, or DNA from cell lysates. Simply add the specially formulated DNA Binding Buffer to the sample and transfer to the supplied column. There is no need for organic denaturants or chloroform. Instead, the product features Fast-Spin column technology to yield high quality, purified DNA in just minutes. The kit quotes high quality purified DNA with a size limit of 73 bp – 23 kb with 25ug of total DNA recovery which can be eluted as little as 25 ul of water.

According to the suggested protocol, before starting, 24 ml 100% ethanol has to be added to the 6 ml DNA Wash Buffer concentrate in order to obtain the final DNA Wash Buffer. The steps which follow are:
1. In a 1.5 ml microcentrifuge tube, add 2 volumes of DNA Binding Buffer to each volume of DNA sample. (e.g., 300 μl binding buffer to 150 μl DNA sample). Mix briefly by vortexing.
2. Transfer mixture to a provided Zymo-Spin™ Column in a Collection Tube.
3. Centrifuge at ≥10,000 x g for 30 seconds. Discard the flow-through.
4. Add 200 μl Wash Buffer to the column. Centrifuge at ≥10,000 x g for 30 seconds. Repeat wash step.
5. Add 30 - 50 μl water or elution buffer directly to the column matrix. Transfer the column to a 1.5 ml microcentrifuge tube and centrifuge at ≥10,000 x g for 30 seconds to elute the DNA.

Ultra-pure DNA in water is now ready for use.

In our laboratory, we use this kit very often for downstream applications like post PCR clean up and post RT clean up of cDNA. The applications we used out of this kit are just 40% of its total applications. This kit can also be used to clean up DNA after any number of enzymatic reactions (e.g. ligase, kinase, phosphatase and more), to purify plasmid DNA from cell-free lysates, for isotope and dye removal or for the isolation of M13 ssDNA.

In our routine applications, we start with 20 ul of sample volume and elute in 30 ul; we have noticed the recovery (in terms of %) goes down if we elute in small volume (i.e. 6 – 10 ul). We have also observed the final yield can be further improved by eluting the DNA with 60 – 70°C water. One of our major downstream applications is gene expression analysis using real-time PCR; we have used this kit to process our samples and have been very satisfied with the results.

I feel the DNA Clean and Concentrator Kit is a perfect choice for efficient DNA clean up which can translate to good, reproducible results in several downstream applications. It is also nice to note that the columns can be bought separately.