HSB

 

 

HSB (f+)

high SDS hybridization buffer (store the main stock at 4oC, working - NT)

Conc.Stock500ml
SDS7%solid35g
Formamid50%100%250ml
283.3g

SSC5x20x125ml
145.3g

Bloc. Reagent2%solid10g
NaP (pH 7.0)50mM1M25ml
27.6g

N-Lauroylsarcosine Na0.1%10%5ml
5.1g

Salm.Sperm DNA50µg/ml10µg/µl2.5ml
H2O mQ49ml

p=1.111

  • Blocking Reagent - is the powder from "Roche Diagnostics GmBH; former Boehringer Mannheim" #1096176. Instead of 2% Blocking Reagent one may use 10x Denhardt or 2% non-fat dry milk.
  • it is possible to dissolve the Blocking Reagent only in high strength buffer (in this particular case in H2O+SSC+NaP).
  • Salmon sperm DNA should be shredded and denaturated.