Proteinase K is a mild detergent. It’s a protein extracting and denaturing solution that works best under low-salt conditions. Let’s learn how to make Proteinase K solution from powder using acetic acid and sterile water..
In A Clean Container, Add 100 Ml of Sterlie Water.
Proteinase K is a proteolytic enzyme that is used in molecular biology for the digestion of proteins. It is also known as proteinase 3, and is found in several species of bacteria, though often as an inactive form. The active form can be extracted from different species of bacteria, or produced synthetically. In order to be active, Proteinase K must be dissolved in water at a sufficient concentration and temperature. The enzyme cannot be stored for long periods of time because it loses activity over time.
The purpose of this step-by-step guide to preparing proteinase K solution is to provide a resource for those who wish to prepare their own proteinase K solution. This guide will show you how to produce your own 100 µg/ml (1mg/ml) solution from powder, as well as how to make a 1% stock solution if you would prefer not to keep large amounts of the enzyme on hand.
Preparation of a 1% stock solution: In a clean container, add 100 ml of sterile water. Add 1 g of Proteinase K powder and mix thoroughly by shaking or stirring for about 2 minutes. Label the container “1% stock solution”. You may now use this solution immediately, or you may store it.
Add The Entire Bottle of Proteinase K Powder to The Water.
Proteinase K, a protein-digesting enzyme, is used in many different applications such as DNA extraction. However, it is also useful in the preparation of samples for other downstream applications such as PCR and TAE (Tris-acetate-EDTA). In order to do this, you need to make a Proteinase K solution from powder. This can be done by following the steps below:
1) In a clean container, add 100 ml of sterlie water. Add the entire bottle of Proteinase K powder to the water.
2) Stir with a stirbar until all of the powder has dissolved.
3) Let the solution sit for about 1 hour before use.
Resuspend The Powder with A Pipette.
Proteinase K is sold as a powder. As a powder, it falls out of solution very easily and needs to be resuspended regularly to keep it from clumping. Here’s how you can make your own Proteinase K solution:
1. Use a micropipette or other fine-tipped pipette to transfer the powder into a glass vial.
2. Add deionized water to the vial until full, being careful not to get any air bubbles into the vial.
3. Place the vial in a water bath that is set between 65 and 75 degrees Celsius for 12 hours. The heat will dissolve the proteinase K.
Shake The Solution.
Before you can use your home-made Proteinase K solution, you need to make sure that it has been thoroughly mixed. You do this by shaking the bottle vigorously and holding it upside down for a few moments. This ensures that the powder and water are evenly combined and there are no clumps of powder left in the bottom of the bottle.
Allow Proteinase K Solution to Sit at Room Temperature to Dissolve Entirely.
Proteinase K Solution is a biological reagent used to prepare tissue samples for RNA isolation. It is commonly sold as a powder which must be dissolved in water before use. However, the powder may not completely dissolve in solutions that contain less than 30% glycerol or are too acidic or basic.
For best results, prepare Proteinase K Solution by dissolving approximately 15 grams of concentrate in 250 mL of DI water and allowing the solution to sit at room temperature (15-25°C) for at least several hours.
Shake Occasionally Over Several Hours Until the Powder Is Dissolved.
If you’ve been reading a lot of the material I’ve written on my blog, you’ve seen me mention the need to make a proteinase K solution to help in recovering lost data. It’s a relatively simple process that takes almost no time at all, but I have seen some people have problems with it because they didn’t shake the solution enough over a long period of time.
So here’s how you do it:
1) Take an old, used glass bottle and fill it up with enough water to fully dissolve the powder inside.
2) Add one scoop of proteinase K powder and stir until it dissolves.
3) Let it sit for several hours and shake it occasionally.
Once The Proteinase K Solution Is Completely Dissolved, Aliquot Iinto Sterile 0.5ml Tubes, Cap And Store At -20C.
Proteinase K solution is used to digest proteins in PCR reaction mixtures. It is made by mixing 3 parts of 80% ethanol, 1 part of 10x phosphatase buffer, and 0.1 part (1 microliter) of proteinase K powder. The mixture is vortexed until it is completely dissolved into a clear solution, then aliquoted and frozen.
Last Words
At the moment, BlenderBottle is the only manufacturer that produces Proteinase K Solution Pre-Mixed.