1 2 3 4

10X PBS (2L)

175.3gNaCl 7.7gKH2PO4 25gK2HPO4 (H2O to 2L)

4% PFA in PBS (50 mls):

50 ml PBS (heated to 60ºC, stirring) 2 g paraformaldehyde .25 ml NaOH .25 ml HCl (add when PFA dissolved) 99.9 % PBS (500 ml) 0.1% Tween 20 (.5 ml) 6% H202 (10 ml 30% H2O2) 94% PBT (40 ml) 50 mls PBT 5 µl 20 mg/ml Proteinase K 50 ml PBT 100 mg glycine 48.75 ml 4% PFA 1.25 ml 8% gluteraldehyde 0.4 ml 25% gluteraldehyde 50%formamide (25 ml) 5X SSC (12.5 ml 20X SSC) 1% SDS(2.5 ml 20% SDS) 50 µg/ml total yeast RNA (250 µl of stock 10 mg/ml tRNA) 50 µg/ml heparin (.87 ml of 5% stock) 50% formamide (250 ml) 5X SSC (125 ml 20X SSC) 1% SDS (25 ml 20% SDS) 50% formamide (250 ml) 2XSSC (50 ml 20X SSC) 80 g NaCl 2 g KCl 25 ml 1 M Tris, pH 7.5 10 ml Tween-20

PBT (500 ml):

6% hydrogen peroxide in PBT (50 mL):

2 mg/ml Proteinase K solution(50 mls):

2 mg/ml glycine in PBT (50 mls):

4% PFA/0.2% gluteraldehyde

or

prehybridization buffer (50 mls):

Solution 1 (500 ml):

Solution 3 (500 ml):

10X TBST(1 l):