Diluent B

Submitted by kdorfman on Mon, 09/26/2022 - 20:21

Dilution buffer for NEB enzymes

Buffer composition:

Reagent Final concentration
NaCl 300 mM
Tris-HCl 10 mM
DTT 1 mM
EDTA 0.1 mM
BSA 500 ug/mL
glycerol 50%
pH 7.4

To make 50 mL for use, make 25 mL of 2x, then mix 1:1 with glycerol

25 mL 2x Diluent B

Reagent c1 c2 v1
NaCl 5M 0.3M 3 mL
Tris HCl 1M 0.01M 0.5 mL
DTT 1 M 1 mM 0.05 mL
EDTA 500 mM 0.1 mM 0.01 mL

plus BSA 25 mg

pH to 7.4, bring volume to 25, add glycerol to 50.

Filter sterilize

Diluent B info sheet from NEB