Proteinase K is a proteolytic enzyme. The enzyme uses Ca2+ ions for structural stability. Due to the high total activity of the enzyme, lysis can be carried out in the presence of EDTA. Proteinase K is therefore excellently usefull for the extraction of nucleic acids from various biological samples.
Recommended Use / Scope
Minerva Biolabs VenorGeM Sample Preparation Kit is used for the extraction of DNA from ordinary samples, like cell culture supernatants containing up to 106 cells, with high efficiency. Anyhow, other samples matrices may contain protein with a concentration above 10 mg/ml, like cell pellets, serum, or fermentation products. These samples may require an additional Proteinase K treatment to increase the overall DNA yield for highest sensitivity.
In combination with Minerva Biolabs´ VenorGeM Sample Preparation Kit 10 µl of the Proteinase K solution is used for each sample after mixing the sample with Conditioner (VenorGeM Sample Preparation Kit) or Buffer A (Microsart AMP Extraction).
All lots of this product are routinely checked by the extraction of 10 CFU/ml of mycoplasma from a defined complex sample matrix in comparison to an untreated sample. The extracts were used as template for mycoplasma PCR with the Microsart® ATMP Mycoplasma Kit for qPCR using a protocol for 10 µl sample volume.
1 vial freeze-dried, for 50 extractions, to be resolved in 500 µl of the included rehydration buffer providing a concentration of 4 mg/ml.
Shelf Life and Storage
The freeze-dried product shall be stored at 2 to 8 °C. The rehydrated stock solution must be stored at < -18 °C. Avoid repeated freezing and thawing. Preparation of aliquots might be required if more than 5 freezing/thawing cycles are required.