mRNA原位杂交步骤

mRNA:1.RNase2.DEPC(0.1%DEPC,3-4,DEPC)3.RNase1.DEPC-PBS(DEPC):140mMNaCL;2.7mMKCL;10mMNa2HPO4;1.8mMKH2PO4pH7.4(DEPC)2.TE:10mMTris,1mMEDTA,pH8.03.0.1M(pH8.0):5.33mlDEPC1L4.20SSC175.3g;88.2gDEPC1L5.100Denhardt’s:Ficoll4002g;PVP()2g;BSA()2g;DEPC100ml6.:20ml(20SSC4ml;4g;10ml;PolyA(10mg/ml)0.5ml;ssDNA(10mg/ml)0.5ml;tRNA(10mg/ml)0.5ml;1M(DTT)2ml;50Denhardt’s0.2ml(-20377.A:100mMTris-HCL(pH7.5);150mMNaCL8.9.10.:A0.1%Triton-X1002%B:100mMTris-HCL(pH9.5);100mMNaCL;50mMMgCL2C:10mMTris-HCL(pH8.1);1mMEDTA1.,,DEPCddH2O,DEPC-PBS(DEPC)252.DEPC-PBS0.3%Triton-X100153.DEPC-PBS254.TERnase-freeK(K5-20g/ml)3710-300.2MHCL0.1%(100-500g/ml)3720-30()5.,PBS+2mg/ml301,DEPC-PBS25;,DEPC-PBS256.DEPC-PBS4%10-157.DEPC-PBS258.0.1M0.25%10(0.1M0.25%)9.2SSC310.37211.2SSC512.371813.1SSC,1SSC55215,0.5SSC55215,0.5SSC1014.A21015.3016.2-4()17.A21018.B1019.BCIP/NBT2-24(RNA,-BCIP/NBT,-DAB)20.C,21.(),(water-solublemountingmedia),.