NameDescriptionHow to MakeStorage
40 % Acrylamide38 % Acrylamide190 g Acrylamide4oC
(500 ml)2 % Bisacrylamide10 g BisacrylamideDark Bottle
Filter through Whatman #1
NH4OAC7.5 M NH4OAc57.87 g NH4OAcRT
(100 ml)Adjust pH to 7.5 Plastic
(with acetic acid)
Bromocresol50% Glycerol25 ml Glycerol
Green Stop30 mM NaOH0.15 ml of 10 M NaOH
Mix1mM Na2EDTA0.1 ml of 500 mM Na2EDTA
0.25% Bromocresol0.125 g Bromocresol Green
Green
100 X Denhardts2 % PVP 360,00020g PVP-20oC
(1 L)2 % Ficoll 400,00020 g Ficoll type 400(aliquots)
2% BSA20 g BSA (fraction V)
Electrophoresis50 % Glycerol50 ml GlycerolRT
Tracking dye0.25 % Bromphenol0.25 g Bromphenol BluePlastic
(100 ml)Blue
50 mM EDTA5 ml 0.5 M EDTA pH 8.0
45 ml ddH2O
Ethidium10 mg/ml EtBr1 g EthBr4oC
BromideDark Bottle
(100 ml)
10X Hoaglands1 mM NH4H2PO420 ml 0.5 M
(1 L)4 mM KNO380 ml 0.5 M
4 mM Ca(NO3)2-4H2O80 ml 0.5 M
2 mM MgSO440 ml 0.5 M
1 mM NH4NO320 ml 0.5 M
30 mg EDTA3 ml 10 g/L
(Ferric-sodium salt)
Trace Elements25 ml (see below)
Add to 732 ml H2O, in the order listed, with continual stirring.
Store 10X solution and stock solutions at 4oC.
Hoaglands142 mg Boric acid
Trace Elements182 mg MnCl2-4H2O
(500 ml)22 mg ZnSO4-7H2O
(Levy recipe)8 mg CuSO4-5H2O
2 mg H2MoO4
Store at 4oC in dark bottle.
Labelling0.1% SDS1 ml 10% SDSRT
Stop Dye60 mM EDTA12 ml 0.5 M EDTA
(100 ml)0.5% Bromphenol Blue0.5 g
1.5% Blue Dextran1.5 g
10 X MOPS200 mM MOPS167.44 g MOPS free acid4oC
(4 L)50 mM NaOAc27.2 gNaOAcDark Bottle
10 mM Na2EDTA14.88 g Na2EDTA.2H2O
Adjust pH to 7.0
(w/NaOH)
Phenol:CHCl350% TE sat'd Phenol500 ml TE sat'd Phenol4oC
50% CHCl3500 ml CHCl3Dark Bottle
0.1% 8-Hydroxy-0.5 g 8-Hydroxyquinoline
Fill the container of phenol with TE pH 8.0. Place container at 68oC to allow the phenol to dissolve. Remove most of the displaced TE and then add the 8-Hydroxyquinoline and CHCl3. Note: Phenol is from BDH, AnalaR grade.
Prehybridization6 X SSPE1200 ml 20X SSPERT
Hybridization5 X Denhardt's200 ml 100X Denhardt'sPlastic
Solution0.5% SDS200 ml 10% SDS
(4 L)
Salmon sperm 5 mg/mlsuspend in 0.1 M NaCl and stir until dissolved
DNAblend in small blender and autoclave
immediately aliquot to 1.5 ml tubes; freeze at -20oC
Southern0.4 M NaOH64 g NaOHRT
Denaturant0.8 M NaCl187 g NaClPlastic
(4L)
Southern1.5 M NaCl346.6 g NaClRT
Neutralization0.5 M Tris242.2 g Trizma BasePlastic
Buffer (pH 7.6)Add ~130ml conc. HCl
(4L)
20 X SSPE3.6 M NaCl840 g NaClRT
(4L)0.2 M NaH2PO4110 g NaH2PO4Plastic
22 mM EDTA33 g EDTA (tetrasodium salt)
pH = 7.4 (~ 68ml 10N NaOH)
2 X SSPE Wash2 X SSPE400 ml 20 X SSPERT
Solution0.1 % SDS4 g SDSPlastic
(4L)0.1 % Sodium PPi4 g Sodium pyrophosphate
0.2 X SSPE Wash0.2 X SSPE40 ml 20 X SSPERT
Solution0.1 % SDS4 g SDSPlastic
(4L)0.1 % PPi4 g Sodium pyrophosphate
SM buffer5.8 g NaClAutoclave
(1 L)2 g MgSO4.7HOH
(0.96 g anhydrous)
5 ml 2% gelatin
50 ml 1 M Tris-HCl pH 7.5
5 X TBE216 g TRIZMA BaseRT
(4 L)110 g Boric AcidPlastic
8.3 g Na4EDTA
pH = 8 (~13ml Conc. HC1)
100 X TNE1.0 M Tris121.1 g Trizma Base4oC
(1 L)1.0 M NaCl58.44 g NaClPlastic
0.1 M EDTA38.2 g Na4EDTA.2 H2O
pH = 7 (w/ HCl)
10 X TAE400 mM Tris193.76 g Trizma BaseRT
(4 L)22 mM Na2EDTA32.76 g Na2EDTA.2HOHPlastic
pH = 8 (~47ml Glacial Acetic
Acid)
100 X TE1 M Tris121.1 g Trizma BaseRT
(1 L)100 mM EDTA 38.02 g Na4EDTA.2 H2OPlastic
pH to 8.0 (w/ conc HCl)
20 X TELS200 mM Tris24.22 g Trizma BaseRT
(1 L)2 mM Na2EDTA0.75 g Na2EDTA.2 H2OPlastic
4 % SDS40 g SDS
pH = 7.6 (w/ conc HCl)
1.0 M Tris1.0 M Tris/HCl121.1 g Trizma BaseRT
(1 L)pH = (usually7-8; Conc. HCl)Plastic
ddHOH to 1L
Tris-Sucrose50 mM Tris-HCl6.06 g Trizma BaseAutoclave
Buffer25 % Sucrose250 g Sucrose
(1L)10 mM NA2EDTA3.72 g NA2EDTA
Adjuste pH to 8.0
(with conc. HCl)l
Triton Lysis70 mM Tris-HCl7 ml 1M Tris-Cl pH 8.04oC
Buffer70 mM EDTA14 ml 0.5 M EDTA pH 8.0Glass
(100 ml)70 mM EGTA14 ml 0.5 M EGTA pH 8.0Autoclave
0.2 % Triton X-1000.2 ml Triton X-100