In situ checklist (Conlon lab)

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Protocol checklist submitted by VGP from Conlon lab protocols [1]


link to protocol checklist word document [2]



Date:__________ Species:_________ Probes:______________________________


Day 1


0.5 vol DEPC 5min RT ___ ___ ___

PBST 5min RT ___ ___ ___ ___ ___

Proteinase K 15min RT ___

*De-vitellinization 10min ___

PBST 5min RT ___ ___ ___

Triethanolamine 5min RT ___ ___(4ml)

Acetic Acid Anhydride 5min RT ___ ___

PBST 5min RT ___ ___

4% PFA 20min RT ___

PBST 5min RT ___ ___ ___ ___ ___

20%Hyb/80%PBST 5min RT ___

Hyb Buffer o/n 60 ___


Day 2


Probe(s):



Day 3


Hyb Buffer 10min 60 ___

2xSSC 20min 60 ___ ___ ___ ___

0.2xSSC 60min 60 ___ repeat if necessary ___

1xMAB 15min 60 ___

1xMAB 15min RT ___

1xMAB/2%BMBR 60min RT ___

Blocking Buffer 60min RT ___

Antibody o/n 4 ___


Day 4


1xMAB 60min RT ___ ___ ___ ___

1xMAB o/n RT ___


Day 5


AP Buffer 5min RT ___ ___

BM Purple AP Substrate 1-8hrs RT/4 ___

PBS 5min RT ___ ___ ___

Fix:_________________ 30min RT ___ ___

MeOH 60min RT ___ (stored ___)



Day 6


Bleaching _____ RT/4