In an attempt to get a water sample that has higher copy numbers for use in the withering syndrome qPCR assay validation procedures, I isolated gDNA from two water filters from the “big reds” abalone tank. Water filters were numbered 4 and 5, with no other information.
Filters were cut into ~13 strips and processed using the Qiagen DNeasy Blood & Tissue Kit. The filters were mixed with 400uL of Buffer AL and 40uL of Proteinase K (both volumes are double what is recommended in the Qiagen protocol). Samples were vortexed and incubated O/N at 56C. After incubation, 400uL of 100% EtOH was added to each sample (this is double the volume recommended by Qiagen). The manufacturer’s protocol was followed for the remainder. Samples were eluted with 100uL of Buffer AE.