【转帖】试剂配方:DNA操作试剂

SouthernBlotHybridizationBuffer(Saghai,sLab)
Final conc. StockVol.
5×SSC 20×250ml
50mMPB(pH6.8) 0.5M100ml
5×Denhardt’s 50×100ml
2.5mMEDTA (pH8.0)0.5M5ml
100μg/ml ssDNA 5mg/ml 20ml
0.4%SDS 20% 20ml
Dextransulfate 50g
ddH2Oto 1000ml
(Placeabeakeronastirrer,addthesesolutionintheorderofappearanceonebyone.SDSshouldbetheverylastitem.)

WashingoffProbeforRe-hybridizationofBlots(I)
Washingtime: 10min
Finalconc. Stock Vol.
0.1×SSC 20×SSC 20ml
0.1%SDS 10%SDS 40ml
dH2Oto 4000ml

WashingoffProbeforRe-hybridizationofBlots(II)
Washingtime:3min
Finalconc. Stock Vol.
0.1NNaOH 10NNaOH 40ml
0.2%SDS 10%SDS 80ml
dH2Oto 4000ml

WashingoffProbeforRe-hybridizationofBlots(Ⅲ)
Washingtime:20min
Finalconc. Stock Vol.
0.2MTris.(pH7.5) 1MTris.(pH7.5) 800ml
0.1×SSC 20×SSC 20ml
0.2%SDS 10%SDS 80ml
dH2Oto 4000ml

BlueJuice
Finalconc. Stock Vol. Vol.
70%Glycerol 100% 35ml 70ml
0.5×TBE 5× 5ml 10ml
0.2%SDS 10% 1ml 2ml
20mMEDTA 0.5M 2ml 4ml
5mg/mlBromphenolBlue 0.25g 0.5g
5mg/mlXylenecyanol 0.25g 0.5g
dH2Oto 50ml 100ml

EB(10mg/ml)
ehidiumbromide 1g
dH2Oto 100ml
Stironamagneticstirrerforseveralhours.Transferthesolutionto
adarkbottleandstoreat4℃.
Theconcentrationofworksolution:0.5μg/μl(50μlstocksolution
In1000mldH2O).

DecontaminationofEB
ReducetheconcentrationofEB4,mixcarefullythenadd1volumeof2.5NHCl,
mixcarefullyandallowthesolutiontostandatroomtemperature
forseveralhours.Add1volumeof2.5NNaOH,mixanddiscard.