Hello,
I am searching for a tris-lysis buffer for the DNA extraction from Artemia (brine shrimp). Does anybody have a lysis buffer solution they can share?
I found the following, but missing pH values:
75mM NaCl
10mM Tris-HCl (ph?)
0.5mM EDTA (pH?)
0.5% SDS
Can somebody confirm that this is correct? Does somebody have a similar solution they are using?
I am using the DNA for MinION sequencing, so I need a lot of HMW (6+ ng), pure DNA.
Thank you!
Procedure to prepare a lysis solution containing 10mM Tris-HCl buffer, 1mM EDTA as the chelating agent, and 0.5% SDS as the detergent.
Step 1: Preparation of 1 L of 1 M Tris-HCl (pH 8) stock solution
Dissolve 121 g Tris-HCL (molecular weight = 157.60 g) in 800 ml distilled water, adjust the pH to 8 using HCl solution, and make up the volume to 1 L using distilled water.
Step 2: Preparation of 500 ml of 0.5 M EDTA stock solution
Dissolve 93.0 g of EDTA [EDTA.Na2.2H2O] (molecular weight = 372.24 g)] in 400 ml of distilled water, add 10 g (approx.) NaOH pellet to adjust pH to 8, and make up the volume to 500 ml using distilled water.
Step 3: Preparation of 10% SDS stock solution
Dissolve 10 g of SDS in 90 ml distilled water, and make up the volume to 100 ml using distilled water.
Step 4: Preparation of 500 ml of the Tris-EDTA SDS lysis buffer
Add 5 ml of 1 M Tris-HCl (pH 8), 1 ml 0.5 M EDTA, and 5 ml of 10% SDS solution to 400 ml of distilled water. Make up the volume to 500 ml.
Procedure to prepare a lysis solution containing 10mM Tris-HCl buffer, 1mM EDTA as the chelating agent, and 0.5% SDS as the detergent.
Step 1: Preparation of 1 L of 1 M Tris-HCl (pH 8) stock solution
Dissolve 121 g Tris-HCL (molecular weight = 157.60 g) in 800 ml distilled water, adjust the pH to 8 using HCl solution, and make up the volume to 1 L using distilled water.
Step 2: Preparation of 500 ml of 0.5 M EDTA stock solution
Dissolve 93.0 g of EDTA [EDTA.Na2.2H2O] (molecular weight = 372.24 g)] in 400 ml of distilled water, add 10 g (approx.) NaOH pellet to adjust pH to 8, and make up the volume to 500 ml using distilled water.
Step 3: Preparation of 10% SDS stock solution
Dissolve 10 g of SDS in 90 ml distilled water, and make up the volume to 100 ml using distilled water.
Step 4: Preparation of 500 ml of the Tris-EDTA SDS lysis buffer
Add 5 ml of 1 M Tris-HCl (pH 8), 1 ml 0.5 M EDTA, and 5 ml of 10% SDS solution to 400 ml of distilled water. Make up the volume to 500 ml.
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