Branchial arch 3 and In situ checklist (Conlon lab): Difference between pages

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(Created page with "Protocol checklist submitted by VGP from Conlon lab protocols [http://www.unc.edu/~fconlon/Protocols.htm#tropprot] link to protocol checklist word document [http://www.unc.edu/...")
 
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#REDIRECT [[XB-ANAT-425]]
Protocol checklist submitted by VGP from Conlon lab protocols [http://www.unc.edu/~fconlon/Protocols.htm#tropprot]
 
 
link to protocol checklist word document [http://www.unc.edu/~fconlon/Protocols/In_Situ_Checklist.doc]
 
 
 
 
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

Latest revision as of 08:52, 18 November 2011

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