Qiagen Vs. Norgen DNA Mini Prep Kits.

OriRi is a difficult origin of replication to qualitatively assay because it is single copy and most DNA prep kits can not isolate enough DNA to visualize the DNA fragments. A Qiagen mini prep kit is currently used for low copy plasmids but is not sensitive enough to use for single copy plasmids and it is hard to see the restriction enzyme digested with the OriRi replication in the Qiagen kit. This study aims to test low copy plasmids with OriV and single copy plasmids with OriRi origins of replication isolated from Agrobacterium with a new DNA prep kit called Norgen Low Copy Plamisd Miniprep kit. The kit is supplied by The Norgen Biotek Corporation  and claimed to designed for low copy plasmid preparation and generating higher quality DNA preparations for microfluidic quality checks. Presently, we are using phi29 rolling circle amplification to allow us to visualize bands. This adds over 18 hours to the QC process and is very expensive, so we hope it will be better in the new kit. Norgen states low copy plasmids are purified based on a spin column chromatopography using Norgen’s proprietary resin as the separation matrix. The DNA is binded under high salt concentrations then releases the bound DNA under low salt and lightly alkali. Low Copy Plamisd Miniprep kit is tested to find out if it would generate higher quality DNA than the control Qiagen kit for low copy plasmids and can isolate enough DNA from single copy plasmids to be visible with microfluidic screenings [This work was supported by National Science Foundation, HBCU-UP awarded to Dr. Sarwan Dhir at Fort Valley State University].