Lysis buffer I
Preparation
Lysis buffer I
Step 1.
Prepare ingredients.
1M Tris (pH 8)
0.5
mL
(10 mM from 1 M stock)
5M NaCl Stock
1
mL
(100 mM from 5 M stock)
0.5M EDTA STOCK
2.5
mL
(25 mM from 0.5 M stock)
10% SDS
2.5
mL
(0.5 % w/v from 10 % w/v stock)
Step 2.
Bring final volume to 50 mL with water.
Nested recipe inputs
Used in step 1
1M Tris (pH 8)
Step 1.
Weigh out the Tris and add to a media bottle.
Tris Base
12.114
g
Step 2.
Fill media bottle to with ~80% target volume with distilled water.
Step 3.
Add a magnetic stirrer and place on a stirring plate to mix the solution.
Step 4.
Use a pH meter to measure pH. Slowly add concentrated hydrochloric acid (HCl) solution using a Pasteur pipette to reduce the pH to 8.0.
Be careful not to add too much at a time, since the pH will change rapidly.
Step 5.
Once the desired pH has been reached, bring to final volume using distilled water.
Step 6.
Bring final volume to 100 mL with water.
Used in step 1
5M NaCl Stock
Step 1.
Prepare ingredients.
Sodium Chloride
14.61
g
Step 2.
Bring final volume to 50 mL with water.
Used in step 1
0.5M EDTA STOCK
Step 1.
Prepare ingredients.
EDTA (Ethylenediaminetetraacetic acid)
7.306
g
Step 2.
Bring final volume to 50 mL with water.
Used in step 1
10% SDS
Step 1.
Prepare ingredients.
Sodium Lauryl Sulfate
5
g
Step 2.
Bring final volume to 50 mL with water.